Accelerated cell-sheet recovery from a surface successively grafted with polyacrylamide and poly(N-isopropylacrylamide).

PubMed

Akiyama, Yoshikatsu; Kikuchi, Akihiko; Yamato, Masayuki; Okano, Teruo

2014-08-01

A double polymeric nanolayer consisting of poly(N-isopropylacrylamide) (PIPAAm) and hydrophilic polyacrylamide (PAAm) was deposited on tissue culture polystyrene (TCPS) surfaces using electron beam irradiation to form a new temperature-responsive cell culture surface in which the basal hydrophilic PAAm component in the double polymeric layer promotes the hydration of the upper PIPAAm layer and induces rapid cell detachment compared to a conventional temperature-responsive cell culture surface, PIPAAm-grafted TCPS (PIPAAm-TCPS). Take-off angle-dependent X-ray photoelectron spectroscopy spectral analysis demonstrated that the grafted PIPAAm and PAAm components were located in the upper and basal regions of the double polymeric layer, respectively, suggesting that the double polymeric layer forms an inter-penetrating-network-like structure with PAAm at the basal portion of the PIPAAm grafted chains. The wettability of the temperature-responsive cell culture surfaces with the double polymeric layer tended to be more hydrophilic, with an increase in the basal PAAm graft density at a constant PIPAAm graft density. However, when the graft densities of the upper PIPAAm and basal PAAm were optimized, the resulting temperature-responsive cell culture surface with the double polymeric layer exhibited rapid cell detachment while maintaining cell adhesive character comparable to that of PIPAAm-TCPS. The cell adhesive character was altered from cell-adhesive to cell-repellent with increasing PAAm or PIPAAm graft density. The cell adhesive character of the temperature-responsive cell culture surfaces was relatively consistent with their contact angles. These results strongly suggest that the basal PAAm surface properties affect the degree of hydration and dehydration of the subsequently grafted PIPAAm. In addition, the roles of the hydrophilic component in accelerating cell detachment are further discussed in terms of the mobility of the grafted PIPAAm chains. Applications of this insight might be useful for designing temperature-responsive cell culture surfaces for achieving efficient cell culture and quick target cell detachment. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Bacterial adhesion force quantification by fluidic force microscopy

NASA Astrophysics Data System (ADS)

Potthoff, Eva; Ossola, Dario; Zambelli, Tomaso; Vorholt, Julia A.

2015-02-01

Quantification of detachment forces between bacteria and substrates facilitates the understanding of the bacterial adhesion process that affects cell physiology and survival. Here, we present a method that allows for serial, single bacterial cell force spectroscopy by combining the force control of atomic force microscopy with microfluidics. Reversible bacterial cell immobilization under physiological conditions on the pyramidal tip of a microchanneled cantilever is achieved by underpressure. Using the fluidic force microscopy technology (FluidFM), we achieve immobilization forces greater than those of state-of-the-art cell-cantilever binding as demonstrated by the detachment of Escherichia coli from polydopamine with recorded forces between 4 and 8 nN for many cells. The contact time and setpoint dependence of the adhesion forces of E. coli and Streptococcus pyogenes, as well as the sequential detachment of bacteria out of a chain, are shown, revealing distinct force patterns in the detachment curves. This study demonstrates the potential of the FluidFM technology for quantitative bacterial adhesion measurements of cell-substrate and cell-cell interactions that are relevant in biofilms and infection biology.Quantification of detachment forces between bacteria and substrates facilitates the understanding of the bacterial adhesion process that affects cell physiology and survival. Here, we present a method that allows for serial, single bacterial cell force spectroscopy by combining the force control of atomic force microscopy with microfluidics. Reversible bacterial cell immobilization under physiological conditions on the pyramidal tip of a microchanneled cantilever is achieved by underpressure. Using the fluidic force microscopy technology (FluidFM), we achieve immobilization forces greater than those of state-of-the-art cell-cantilever binding as demonstrated by the detachment of Escherichia coli from polydopamine with recorded forces between 4 and 8 nN for many cells. The contact time and setpoint dependence of the adhesion forces of E. coli and Streptococcus pyogenes, as well as the sequential detachment of bacteria out of a chain, are shown, revealing distinct force patterns in the detachment curves. This study demonstrates the potential of the FluidFM technology for quantitative bacterial adhesion measurements of cell-substrate and cell-cell interactions that are relevant in biofilms and infection biology. Electronic supplementary information (ESI) available: Video S1. Detachment of a S. pyogenes cell chain from glass substrate. The cantilever is approached on the outermost adherent cell of a chain and four bacteria were then sequentially detached. The sequential cell detachment suddenly stopped after four bacteria. This possibly occurred because bacteria-glass interactions became too strong or the maximal probe retraction was reached. The cells spontaneously detached from the cantilever flipping back on the surface. Fig. S1. (A) Adhesion force-distance and (B) adhesion force-detaching work correlation of E.coli on PLL for setpoints of 1 and 10 nN. Circle: 1 nN setpoint, square: 10 nN. See DOI: 10.1039/c4nr06495j

Influence of airway wall compliance on epithelial cell injury and adhesion during interfacial flows

PubMed Central

Higuita-Castro, Natalia; Mihai, Cosmin; Hansford, Derek J.

2014-01-01

Interfacial flows during cyclic airway reopening are an important source of ventilator-induced lung injury. However, it is not known how changes in airway wall compliance influence cell injury during airway reopening. We used an in vitro model of airway reopening in a compliant microchannel to investigate how airway wall stiffness influences epithelial cell injury. Epithelial cells were grown on gel substrates with different rigidities, and cellular responses to substrate stiffness were evaluated in terms of metabolic activity, mechanics, morphology, and adhesion. Repeated microbubble propagations were used to simulate cyclic airway reopening, and cell injury and detachment were quantified via live/dead staining. Although cells cultured on softer gels exhibited a reduced elastic modulus, these cells experienced less plasma membrane rupture/necrosis. Cells on rigid gels exhibited a minor, but statistically significant, increase in the power law exponent and also exhibited a significantly larger height-to-length aspect ratio. Previous studies indicate that this change in morphology amplifies interfacial stresses and, therefore, correlates with the increased necrosis observed during airway reopening. Although cells cultured on stiff substrates exhibited more plasma membrane rupture, these cells experienced significantly less detachment and monolayer disruption during airway reopening. Western blotting and immunofluorescence indicate that this protection from detachment and monolayer disruption correlates with increased focal adhesion kinase and phosphorylated paxillin expression. Therefore, changes in cell morphology and focal adhesion structure may govern injury responses during compliant airway reopening. In addition, these results indicate that changes in airway compliance, as occurs during fibrosis or emphysema, may significantly influence cell injury during mechanical ventilation. PMID:25213636

Modeling cell-substrate de-adhesion dynamics under fluid shear

NASA Astrophysics Data System (ADS)

Maan, Renu; Rani, Garima; Menon, Gautam I.; Pullarkat, Pramod A.

2018-07-01

Changes in cell-substrate adhesion are believed to signal the onset of cancer metastasis, but such changes must be quantified against background levels of intrinsic heterogeneity between cells. Variations in cell-substrate adhesion strengths can be probed through biophysical measurements of cell detachment from substrates upon the application of an external force. Here, we investigate, theoretically and experimentally, the detachment of cells adhered to substrates when these cells are subjected to fluid shear. We present a theoretical framework within which we calculate the fraction of detached cells as a function of shear stress for fast ramps as well as the decay in this fraction at fixed shear stress as a function of time. Using HEK and 3T3 fibroblast cells as experimental model systems, we extract characteristic force scales for cell adhesion as well as characteristic detachment times. We estimate force-scales of  ∼500 pN associated to a single focal contact, and characteristic time-scales of s representing cell-spread-area dependent mean first passage times to the detached state at intermediate values of the shear stress. Variations in adhesion across cell types are especially prominent when cell detachment is probed by applying a time-varying shear stress. These methods can be applied to characterizing changes in cell adhesion in a variety of contexts, including metastasis.

Assessment of the biocompatibility, stability, and suitability of novel thermoresponsive films for the rapid generation of cellular constructs

NASA Astrophysics Data System (ADS)

Reed, Jamie A.

2011-12-01

Stimuli responsive polymers (SRP) are of great interest in the bioengineering community due to their use in applications such as drug delivery and tissue engineering. One example of an SRP is poly(N-isopropyl acrylamide) or pNIPAM. This SRP has the capability of changing its conformation with a slight temperature change: adherent mammalian cells spontaneously release as a confluent cell sheet, which can be harvested for cell sheet engineering purposes. Since its initial use in 1968, many researchers have used pNIPAM to obtain a cell sheet composed of their cell type of interest. The differing protocols used for these diverse cell types, such as the conditions used for cell detachment, and the varying methods used for derivatizing substrates with pNIPAM have all led to conflicting reports on the utility of pNIPAM for cell sheet engineering purposes, as well as the relative cytotoxicity of the polymer. In this work, some of the key inconsistencies in the literature and previously unaddressed challenges when utilizing pNIPAM films are overcome for the purpose of rapid generation of cellular constructs, specifically spheroids. Pertinent characteristics of low temperature detachment are investigated for their effect on the kinetics of cell detachment. In addition, a novel, inexpensive method for obtaining pNIPAM films for mammalian cell detachment, combining pNIPAM with a sol-gel, was optimized and compared to plasma polymerization deposition. Furthermore, proper storage conditions (e.g. temperature and relative humidity) for these films were investigated to increase stability of the films for using tissue culture conditions. To increase the speed of generation of cell sheets, electrospun mats and hydrogels with a high surface area-to-volume ratio were developed. The result is a platform appropriate for the rapid formation of cellular constructs, such as engineered tissues and spheroids for cancer cell research.

Accelerated cell sheet detachment by copolymerizing hydrophilic PEG side chains into PNIPAm nanocomposite hydrogels.

PubMed

Liu, Dan; Wang, Tao; Liu, Xinxing; Tong, Zhen

2012-10-01

One-end-connected short poly(ethylene glycol) (PEG) side chains were facilely introduced into the poly(N-isopropylacrylamide) (PNIPAm) nanocomposite hydrogel (NC gel) via in situ copolymerization of NIPAm monomer and PEG macromonomer in the aqueous suspension of hectorite clay Laponite XLS. The NC gels were characterized with Fourier transform infrared and x-ray photoelectron spectroscopy for the composition, DSC and transmittance for the phase separation temperature, dynamic mechanical spectra and swelling ratio for the interaction. Increasing the PEG content led to a small increase in the storage modulus and the lower critical solution temperature (LCST) of the copolymerized NC gels, and the LCST of the copolymerized NC gels was still below 37 °C. The L929 cell adhesion and proliferation on the surface of these NC gels were not suppressed by the incorporation of hydrophilic PEG side chains. By lowering temperature below the LCST, the cell sheet spontaneously detached from the copolymerized NC gels. The surface morphology and surface wettability of the NC gels were detected by atom force microscope and contact angle measurement. A rough and hydrophilic surface induced by a small amount of PEG side chains was found to be favorable to accelerate the cell sheet detachment, probably due to the enhanced water permeation into the gel-cell sheet interface.

Traction force microscopy in rapidly moving cells reveals separate roles for ROCK and MLCK in the mechanics of retraction.

PubMed

Morin, Timothy R; Ghassem-Zadeh, Sean A; Lee, Juliet

2014-08-15

Retraction is a major rate-limiting step in cell motility, particularly in slow moving cell types that form large stable adhesions. Myosin II dependent contractile forces are thought to facilitate detachment by physically pulling up the rear edge. However, retraction can occur in the absence of myosin II activity in cell types that form small labile adhesions. To investigate the role of contractile force generation in retraction, we performed traction force microscopy during the movement of fish epithelial keratocytes. By correlating changes in local traction stress at the rear with the area retracted, we identified four distinct modes of retraction. "Recoil" retractions are preceded by a rise in local traction stress, while rear edge is temporarily stuck, followed by a sharp drop in traction stress upon detachment. This retraction type was most common in cells generating high average traction stress. In "pull" type retractions local traction stress and area retracted increase concomitantly. This was the predominant type of retraction in keratocytes and was observed mostly in cells generating low average traction stress. "Continuous" type retractions occur without any detectable change in traction stress, and are seen in cells generating low average traction stress. In contrast, to many other cell types, "release" type retractions occur in keratocytes following a decrease in local traction stress. Our identification of distinct modes of retraction suggests that contractile forces may play different roles in detachment that are related to rear adhesion strength. To determine how the regulation of contractility via MLCK or Rho kinase contributes to the mechanics of detachment, inhibitors were used to block or augment these pathways. Modulation of MLCK activity led to the most rapid change in local traction stress suggesting its importance in regulating attachment strength. Surprisingly, Rho kinase was not required for detachment, but was essential for localizing retraction to the rear. We suggest that in keratocytes MLCK and Rho kinase play distinct, complementary roles in the respective temporal and spatial control of rear detachment that is essential for maintaining rapid motility. Copyright © 2014 Elsevier Inc. All rights reserved.

Gelatin promotes murine fibrosarcoma L929 cell detachment and protects the cells from TNFα-induced cytotoxicity.

PubMed

Wang, Hong-Ju; Li, Meng-Qi; Liu, Wei; Yao, Guo-Dong; Xia, Ming-Yu; Hayashi, Toshihiko; Fujisaki, Hitomi; Hattori, Shunji; Tashiro, Shin-Ichi; Onodera, Satoshi; Ikejima, Takashi

2016-07-01

Gelatin has been considered to exist as intermediate substance of collagen catabolism in tissue remodeling or under inflammatory conditions. We have initiated the study on possible biological functions of gelatin that can exist temporally and locally under the conditions of remodeling and inflammation Materials and methods: To this purpose, we investigated cell proliferation and survival on gelatin-coated dishes and the response to tumor necrosis factor α (TNFα)-induced cytotoxicity in L929 cells. Autophagy level, ATP level, and ROS generation are examined. L929 cells detached from the gelatin-coated dishes and formed multicellular aggregates. TNFα-induced cytotoxicity in L929 cells was inhibited by gelatin-coating culture. The cells on gelatin-coated dishes showed reduced cellular ATP levels and increased adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) phosphorylation, leading to increased ROS generation and autophagy. This study showed that gelatin-coated culture protected L929 cells from TNFα-induced cytotoxicity and suggested for a possible pathophysiological function of gelatin in regulating cellular functions.

Fibronectin matrix-mediated cohesion suppresses invasion of prostate cancer cells.

PubMed

Jia, Dongxuan; Entersz, Ildiko; Butler, Christine; Foty, Ramsey A

2012-03-20

Invasion is an important early step in the metastatic cascade and is the primary cause of death of prostate cancer patients. In order to invade, cells must detach from the primary tumor. Cell-cell and cell-ECM interactions are important regulators of cohesion--a property previously demonstrated to mediate cell detachment and invasion. The studies reported here propose a novel role for α5β1 integrin--the principle mediator of fibronectin matrix assembly (FNMA)--as an invasion suppressor of prostate cancer cells. Using a combination of biophysical and cell biological methods, and well-characterized prostate cancer cell lines of varying invasiveness, we explore the relationship between cohesion, invasiveness, and FNMA. We show that cohesion is inversely proportional to invasive capacity. We also show that more invasive cells express lower levels of α5β1 integrin and lack the capacity for FNMA. Cells were generated to over-express either wild-type α5 integrin or an integrin in which the cytoplasmic domain of α5 was replaced with that of α2. The α2 construct does not promote FNMA. We show that only wild-type α5 integrin promotes aggregate compaction, increases cohesion, and reduces invasion of the more aggressive cells, and that these effects can be blocked by the 70-kDa fibronectin fragment. We propose that restoring capacity for FNMA in deficient cells can increase tumor intercellular cohesion to a point that significantly reduces cell detachment and subsequent invasion. In prostate cancer, this could be of therapeutic benefit by blocking an early key step in the metastatic cascade.

Cell-Adhesive and Cell-Repulsive Zwitterionic Oligopeptides for Micropatterning and Rapid Electrochemical Detachment of Cells

PubMed Central

Kakegawa, Takahiro; Mochizuki, Naoto; Sadr, Nasser; Suzuki, Hiroaki

2013-01-01

In this study, we describe the development of oligopeptide-modified cell culture surfaces from which adherent cells can be rapidly detached by application of an electrical stimulus. An oligopeptide, CGGGKEKEKEK, was designed with a terminal cysteine residue to mediate binding to a gold surface via a gold–thiolate bond. The peptide forms a self-assembled monolayer through the electrostatic force between the sequence of alternating charged glutamic acid (E) and lysine (K) residues. The dense and electrically neutral oligopeptide zwitterionic layer of the modified surface was resistant to nonspecific adsorption of proteins and adhesion of cells, while the surface was altered to cell adhesive by the addition of a second oligopeptide (CGGGKEKEKEKGRGDSP) containing the RGD cell adhesion motif. Application of a negative electrical potential to this gold surface cleaved the gold–thiolate bond, leading to desorption of the oligopeptide layer, and rapid (within 2 min) detachment of virtually all cells. This approach was applicable not only to detachment of cell sheets but also for transfer of cell micropatterns to a hydrogel. This electrochemical approach of cell detachment may be a useful tool for tissue-engineering applications. PMID:22853640

Lipid tethering of breast tumor cells enables real-time imaging of free-floating cell dynamics and drug response

PubMed Central

Whipple, Rebecca A.; Zhang, Peipei; Sooklal, Elisabeth L.; Martin, Stuart S.; Jewell, Christopher M.

2016-01-01

Free-floating tumor cells located in the blood of cancer patients, known as circulating tumor cells (CTCs), have become key targets for studying metastasis. However, effective strategies to study the free-floating behavior of tumor cells in vitro have been a major barrier limiting the understanding of the functional properties of CTCs. Upon extracellular-matrix (ECM) detachment, breast tumor cells form tubulin-based protrusions known as microtentacles (McTNs) that play a role in the aggregation and re-attachment of tumor cells to increase their metastatic efficiency. In this study, we have designed a strategy to spatially immobilize ECM-detached tumor cells while maintaining their free-floating character. We use polyelectrolyte multilayers deposited on microfluidic substrates to prevent tumor cell adhesion and the addition of lipid moieties to tether tumor cells to these surfaces through interactions with the cell membranes. This coating remains optically clear, allowing capture of high-resolution images and videos of McTNs on viable free-floating cells. In addition, we show that tethering allows for the real-time analysis of McTN dynamics on individual tumor cells and in response to tubulin-targeting drugs. The ability to image detached tumor cells can vastly enhance our understanding of CTCs under conditions that better recapitulate the microenvironments they encounter during metastasis. PMID:26871289

Analysis of Bacterial Detachment from Substratum Surfaces by the Passage of Air-Liquid Interfaces

PubMed Central

Gómez-Suárez, Cristina; Busscher, Henk J.; van der Mei, Henny C.

2001-01-01

A theoretical analysis of the detachment of bacteria adhering to substratum surfaces upon the passage of an air-liquid interface is given, together with experimental results for bacterial detachment in the absence and presence of a conditioning film on different substratum surfaces. Bacteria (Streptococcus sobrinus HG1025, Streptococcus oralis J22, Actinomyces naeslundii T14V-J1, Bacteroides fragilis 793E, and Pseudomonas aeruginosa 974K) were first allowed to adhere to hydrophilic glass and hydrophobic dimethyldichlorosilane (DDS)-coated glass in a parallel-plate flow chamber until a density of 4 × 106 cells cm−2 was reached. For S. sobrinus HG1025, S. oralis J22, and A. naeslundii T14V-J1, the conditioning film consisted of adsorbed salivary components, while for B. fragilis 793E and P. aeruginosa 974K, the film consisted of adsorbed human plasma components. Subsequently, air bubbles were passed through the flow chamber and the bacterial detachment percentages were measured. For some experimental conditions, like with P. aeruginosa 974K adhering to DDS-coated glass and an air bubble moving at high velocity (i.e., 13.6 mm s−1), no bacteria detached upon passage of an air-liquid interface, while for others, detachment percentages between 80 and 90% were observed. The detachment percentage increased when the velocity of the passing air bubble decreased, regardless of the bacterial strain and substratum surface hydrophobicity involved. However, the variation in percentages of detachment by a passing air bubble depended greatly upon the strain and substratum surface involved. At low air bubble velocities the hydrophobicity of the substratum had no influence on the detachment, but at high air bubble velocities all bacterial strains were more efficiently detached from hydrophilic glass substrata. Furthermore, the presence of a conditioning film could either inhibit or stimulate detachment. The shape of the bacterial cell played a major role in detachment at high air bubble velocities, and spherical strains (i.e., streptococci) detached more efficiently than rod-shaped organisms. The present results demonstrate that methodologies to study bacterial adhesion which include contact with a moving air-liquid interface (i.e., rinsing and dipping) yield detachment of an unpredictable number of adhering microorganisms. Hence, results of studies based on such methodologies should be referred as “bacterial retention” rather than “bacterial adhesion”. PMID:11375160

Analysis of bacterial detachment from substratum surfaces by the passage of air-liquid interfaces.

PubMed

Gómez-Suárez, C; Busscher, H J; van der Mei, H C

2001-06-01

A theoretical analysis of the detachment of bacteria adhering to substratum surfaces upon the passage of an air-liquid interface is given, together with experimental results for bacterial detachment in the absence and presence of a conditioning film on different substratum surfaces. Bacteria (Streptococcus sobrinus HG1025, Streptococcus oralis J22, Actinomyces naeslundii T14V-J1, Bacteroides fragilis 793E, and Pseudomonas aeruginosa 974K) were first allowed to adhere to hydrophilic glass and hydrophobic dimethyldichlorosilane (DDS)-coated glass in a parallel-plate flow chamber until a density of 4 x 10(6) cells cm(-2) was reached. For S. sobrinus HG1025, S. oralis J22, and A. naeslundii T14V-J1, the conditioning film consisted of adsorbed salivary components, while for B. fragilis 793E and P. aeruginosa 974K, the film consisted of adsorbed human plasma components. Subsequently, air bubbles were passed through the flow chamber and the bacterial detachment percentages were measured. For some experimental conditions, like with P. aeruginosa 974K adhering to DDS-coated glass and an air bubble moving at high velocity (i.e., 13.6 mm s(-1)), no bacteria detached upon passage of an air-liquid interface, while for others, detachment percentages between 80 and 90% were observed. The detachment percentage increased when the velocity of the passing air bubble decreased, regardless of the bacterial strain and substratum surface hydrophobicity involved. However, the variation in percentages of detachment by a passing air bubble depended greatly upon the strain and substratum surface involved. At low air bubble velocities the hydrophobicity of the substratum had no influence on the detachment, but at high air bubble velocities all bacterial strains were more efficiently detached from hydrophilic glass substrata. Furthermore, the presence of a conditioning film could either inhibit or stimulate detachment. The shape of the bacterial cell played a major role in detachment at high air bubble velocities, and spherical strains (i.e., streptococci) detached more efficiently than rod-shaped organisms. The present results demonstrate that methodologies to study bacterial adhesion which include contact with a moving air-liquid interface (i.e., rinsing and dipping) yield detachment of an unpredictable number of adhering microorganisms. Hence, results of studies based on such methodologies should be referred as "bacterial retention" rather than "bacterial adhesion".

Effectiveness of EDTA and Modified Salt Solution to Detach and Kill Cells from Enterococcus faecalis Biofilm.

PubMed

de Almeida, Josiane; Hoogenkamp, Michel; Felippe, Wilson T; Crielaard, Wim; van der Waal, Suzette V

2016-02-01

Disruption of the matrix of endodontic biofilms will aid in their removal from a root canal. Therefore, the aim of this study was to investigate the efficacy of EDTA and a modified salt solution (MSS) to detach bacteria from biofilms. Forty-eight-hour-old Enterococcus faecalis biofilms were grown on glass coverslips and then treated for 1 hour by immersion in 17% EDTA or MSS. Phosphate-buffered saline served as a negative control. Then, residual biofilm cells on the substrate and the detached cells in the supernatant were collected. Viability was verified by the colony-forming unit (CFU) counting method. Propidium monoazide (PMA) treatment in conjunction with quantitative polymerase chain reaction (qPCR) was also performed to detect the presence of E. faecalis 16S ribonucleic RNA genes. Data were analyzed using 1-way analysis of variance and Tukey or Kruskal-Wallis and Dunn tests. The Pearson R test evaluated the correlation between results from CFU and PMA (α = 5%). qPCR showed that EDTA detached 99% of biofilm cells, and MSS detached 94% of biofilm cells (both P < .001). In contrast to EDTA, MSS was highly antimicrobial. The treatment promoted an ample log 7 reduction of the attached cells (P < .001), and almost no live cells were detected in the supernatant (P < .001). Positive correlations between CFU and qPCR with PMA were observed (r = 0.959 and r = 0.729). EDTA detached cells in biofilms with a minor antimicrobial effect. Besides a great antimicrobial effect, MSS also detached biofilm cells. These dispersals of biofilms give insights into new endodontic biofilm removal strategies. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Evidence that ganglion cells react to retinal detachment.

PubMed

Coblentz, Francie E; Radeke, Monte J; Lewis, Geoffrey P; Fisher, Steven K

2003-03-01

Growth associated protein 43 (GAP 43) is involved in synapse formation and it is expressed in the retina in a very specific pattern. Although GAP 43 is downregulated at the time of synapse formation, it can be re-expressed following injury such as axotomy or ischemia. Because of this we sought to characterize the expression of GAP 43 after retinal detachment (RD). Immunoblot, immunocytochemical and quantitative polymerase chain reaction (QPCR) techniques were used to assess the level of GAP 43 expression after experimental RD. GAP 43 was localized to three sublaminae of the inner plexiform layer of the normal retina. GAP 43 became upregulated in a subset of retinal ganglion cells following at least 7 days of RD. By immunoblot GAP 43 could be detected by 3 days. QPCR shows the upregulation of GAP 43 message by 6hr of detachment. To further characterize changes in ganglion cells, we used an antibody to neurofilament 70 and 200kDa (NF) proteins. Anti-NF labels horizontal cells, ganglion cell dendrites in the inner plexiform layer, and ganglion cell axons (fasicles) in the normal retina. Following detachment it is upregulated in horizontal cells and ganglion cells. When detached retina was double labelled with anti-GAP 43 and anti-NF, some cells were labelled with both markers, while others labelled with only one. We have previously shown that second order neurons respond to detachment; here we show that third order neurons are responding as well. Cellular remodelling of this type in response to detachment may explain the slow recovery of vision that often occurs after reattachment, or those changes that are often assumed to be permanent.

Matrix detachment and proteasomal inhibitors diminish Sulf-2 expression in breast cancer cell lines and mouse xenografts

PubMed Central

Khurana, Ashwani; Jung-Beom, Deok; He, Xiaoping; Kim, Sung-Hoon; Busby, Robert C.; Lorenzon, Laura; Villa, Massimo; Baldi, Alfonso; Molina, Julian; Goetz, Matthew P.; Shridhar, Viji

2013-01-01

Sulfatase 2 (Sulf-2) has been previously shown to be upregulated in breast cancer. Sulf-2 removes sulfate moieties on heparan sulfate proteoglycans which in turn modulate heparin binding growth factor signaling. Here we report that matrix detachment resulted in decreased Sulf-2 expression in breast cancer cells and increased cleavage of poly ADP-ribose polymerase. Silencing of Sulf-2 promotes matrix detachment induced cell death in MCF10DCIS cells. In an attempt to identify Sulf-2 specific inhibitor, we found that proteasomal inhibitors such as MG132, Lactacystin and Bortezomib treatment abolished Sulf-2 expression in multiple breast cancer cell lines. Additionally, we show that Bortezomib treatment of MCF10DCIS cell xenografts in mouse mammary fat pads significantly reduced tumor size, caused massive apoptosis and more importantly reduced Sulf-2 levels in vivo. Finally, our immunohistochemistry analysis of Sulf-2 expression in cohort of patient derived breast tumors indicates that Sulf-2 is significantly upregulated in autologous metastatic lesions compared to primary tumors (p < 0.037, Pearson correlation, Chi-Square analysis). In all, our data suggest that Sulf-2 might play an important role in breast cancer progression from ductal carcinoma in situ into an invasive ductal carcinoma potentially by resisting cell death. PMID:23412907

A Direct Cell Quenching Method for Cell-Culture Based Metabolomics

EPA Science Inventory

A crucial step in metabolomic analysis of cellular extracts is the cell quenching process. The conventional method first uses trypsin to detach cells from their growth surface. This inevitably changes the profile of cellular metabolites since the detachment of cells from the extr...

Quantifying cell adhesion through impingement of a controlled microjet.

PubMed

Visser, Claas Willem; Gielen, Marise V; Hao, Zhenxia; Le Gac, Séverine; Lohse, Detlef; Sun, Chao

2015-01-06

The impingement of a submerged, liquid jet onto a cell-covered surface allows assessing cell attachment on surfaces in a straightforward and quantitative manner and in real time, yielding valuable information on cell adhesion. However, this approach is insufficiently characterized for reliable and routine use. In this work, we both model and measure the shear stress exerted by the jet on the impingement surface in the micrometer-domain, and subsequently correlate this to jet-induced cell detachment. The measured and numerically calculated shear stress data are in good agreement with each other, and with previously published values. Real-time monitoring of the cell detachment reveals the creation of a circular cell-free area upon jet impingement, with two successive detachment regimes: 1), a dynamic regime, during which the cell-free area grows as a function of both the maximum shear stress exerted by the jet and the jet diameter; followed by 2), a stationary regime, with no further evolution of the cell-free area. For the latter regime, which is relevant for cell adhesion strength assessment, a relationship between the jet Reynolds number, the cell-free area, and the cell adhesion strength is proposed. To illustrate the capability of the technique, the adhesion strength of HeLa cervical cancer cells is determined ((34 ± 14) N/m(2)). Real-time visualization of cell detachment in the dynamic regime shows that cells detach either cell-by-cell or by collectively (for which intact parts of the monolayer detach as cell sheets). This process is dictated by the cell monolayer density, with a typical threshold of (1.8 ± 0.2) × 10(9) cells/m(2), above which the collective behavior is mostly observed. The jet impingement method presents great promises for the field of tissue engineering, as the influence of both the shear stress and the surface characteristics on cell adhesion can be systematically studied. Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Nitric Oxide Regulates Lung Carcinoma Cell Anoikis through Inhibition of Ubiquitin-Proteasomal Degradation of Caveolin-1*

PubMed Central

Chanvorachote, Pithi; Nimmannit, Ubonthip; Lu, Yongju; Talbott, Siera; Jiang, Bing-Hua; Rojanasakul, Yon

2009-01-01

Anoikis, a detachment-induced apoptosis, is a principal mechanism of inhibition of tumor cell metastasis. Tumor cells can acquire anoikis resistance which is frequently observed in metastatic lung cancer. This phenomenon becomes an important obstacle of efficient cancer therapy. Recently, signaling mediators such as caveolin-1 (Cav-1) and nitric oxide (NO) have garnered attention in metastasis research; however, their role and the underlying mechanisms of metastasis regulation are largely unknown. Using human lung carcinoma H460 cells, we show that NO impairs the apoptotic function of the cells after detachment. The NO donors sodium nitroprusside and diethylenetriamine NONOate inhibit detachment-induced apoptosis, whereas the NO inhibitors aminoguanidine and 2-(4-carboxyphenyl) tetramethylimidazoline-1-oxyl-3-oxide promote this effect. Resistance to anoikis in H460 cells is mediated by Cav-1, which is significantly down-regulated after cell detachment through a non-transcriptional mechanism involving ubiquitin-proteasomal degradation. NO inhibits this down-regulation by interfering with Cav-1 ubiquitination through a process that involves protein S-nitrosylation, which prevents its proteasomal degradation and induction of anoikis by cell detachment. These findings indicate a novel pathway for NO regulation of Cav-1, which could be a key mechanism of anoikis resistance in tumor cells. PMID:19706615

Nitric oxide regulates lung carcinoma cell anoikis through inhibition of ubiquitin-proteasomal degradation of caveolin-1.

PubMed

Chanvorachote, Pithi; Nimmannit, Ubonthip; Lu, Yongju; Talbott, Siera; Jiang, Bing-Hua; Rojanasakul, Yon

2009-10-09

Anoikis, a detachment-induced apoptosis, is a principal mechanism of inhibition of tumor cell metastasis. Tumor cells can acquire anoikis resistance which is frequently observed in metastatic lung cancer. This phenomenon becomes an important obstacle of efficient cancer therapy. Recently, signaling mediators such as caveolin-1 (Cav-1) and nitric oxide (NO) have garnered attention in metastasis research; however, their role and the underlying mechanisms of metastasis regulation are largely unknown. Using human lung carcinoma H460 cells, we show that NO impairs the apoptotic function of the cells after detachment. The NO donors sodium nitroprusside and diethylenetriamine NONOate inhibit detachment-induced apoptosis, whereas the NO inhibitors aminoguanidine and 2-(4-carboxyphenyl) tetramethylimidazoline-1-oxyl-3-oxide promote this effect. Resistance to anoikis in H460 cells is mediated by Cav-1, which is significantly down-regulated after cell detachment through a non-transcriptional mechanism involving ubiquitin-proteasomal degradation. NO inhibits this down-regulation by interfering with Cav-1 ubiquitination through a process that involves protein S-nitrosylation, which prevents its proteasomal degradation and induction of anoikis by cell detachment. These findings indicate a novel pathway for NO regulation of Cav-1, which could be a key mechanism of anoikis resistance in tumor cells.

Detachment of Chain-Forming Neuroblasts by Fyn-Mediated Control of cell-cell Adhesion in the Postnatal Brain.

PubMed

Fujikake, Kazuma; Sawada, Masato; Hikita, Takao; Seto, Yayoi; Kaneko, Naoko; Herranz-Pérez, Vicente; Dohi, Natsuki; Homma, Natsumi; Osaga, Satoshi; Yanagawa, Yuchio; Akaike, Toshihiro; García-Verdugo, Jose Manuel; Hattori, Mitsuharu; Sobue, Kazuya; Sawamoto, Kazunobu

2018-05-09

In the rodent olfactory system, neuroblasts produced in the ventricular-subventricular zone of the postnatal brain migrate tangentially in chain-like cell aggregates toward the olfactory bulb (OB) through the rostral migratory stream (RMS). After reaching the OB, the chains are dissociated and the neuroblasts migrate individually and radially toward their final destination. The cellular and molecular mechanisms controlling cell-cell adhesion during this detachment remain unclear. Here we report that Fyn, a nonreceptor tyrosine kinase, regulates the detachment of neuroblasts from chains in the male and female mouse OB. By performing chemical screening and in vivo loss-of-function and gain-of-function experiments, we found that Fyn promotes somal disengagement from the chains and is involved in neuronal migration from the RMS into the granule cell layer of the OB. Fyn knockdown or Dab1 (disabled-1) deficiency caused p120-catenin to accumulate and adherens junction-like structures to be sustained at the contact sites between neuroblasts. Moreover, a Fyn and N-cadherin double-knockdown experiment indicated that Fyn regulates the N-cadherin-mediated cell adhesion between neuroblasts. These results suggest that the Fyn-mediated control of cell-cell adhesion is critical for the detachment of chain-forming neuroblasts in the postnatal OB. SIGNIFICANCE STATEMENT In the postnatal brain, newly born neurons (neuroblasts) migrate in chain-like cell aggregates toward their destination, where they are dissociated into individual cells and mature. The cellular and molecular mechanisms controlling the detachment of neuroblasts from chains are not understood. Here we show that Fyn, a nonreceptor tyrosine kinase, promotes the somal detachment of neuroblasts from chains, and that this regulation is critical for the efficient migration of neuroblasts to their destination. We further show that Fyn and Dab1 (disabled-1) decrease the cell-cell adhesion between chain-forming neuroblasts, which involves adherens junction-like structures. Our results suggest that Fyn-mediated regulation of the cell-cell adhesion of neuroblasts is critical for their detachment from chains in the postnatal brain. Copyright © 2018 the authors 0270-6474/18/384599-12$15.00/0.

A single-cell scraper based on an atomic force microscope for detaching a living cell from a substrate

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

Iwata, Futoshi, E-mail: iwata.futoshi@shizuoka.ac.jp; Research Institute of Electronics, Shizuoka University, Johoku, Naka-ku, Hamamatsu 432-8011; Adachi, Makoto

We describe an atomic force microscope (AFM) manipulator that can detach a single, living adhesion cell from its substrate without compromising the cell's viability. The micrometer-scale cell scraper designed for this purpose was fabricated from an AFM micro cantilever using focused ion beam milling. The homemade AFM equipped with the scraper was compact and standalone and could be mounted on a sample stage of an inverted optical microscope. It was possible to move the scraper using selectable modes of operation, either a manual mode with a haptic device or a computer-controlled mode. The viability of the scraped single cells wasmore » evaluated using a fluorescence dye of calcein-acetoxymethl ester. Single cells detached from the substrate were collected by aspiration into a micropipette capillary glass using an electro-osmotic pump. As a demonstration, single HeLa cells were selectively detached from the substrate and collected by the micropipette. It was possible to recultivate HeLa cells from the single cells collected using the system.« less

Thermo-responsive cell culture carriers based on poly(vinyl methyl ether)—the effect of biomolecular ligands to balance cell adhesion and stimulated detachment

NASA Astrophysics Data System (ADS)

Teichmann, Juliane; Nitschke, Mirko; Pette, Dagmar; Valtink, Monika; Gramm, Stefan; Härtel, Frauke V.; Noll, Thomas; Funk, Richard H. W.; Engelmann, Katrin; Werner, Carsten

2015-08-01

Two established material systems for thermally stimulated detachment of adherent cells were combined in a cross-linked polymer blend to merge favorable properties. Through this approach poly(N-isopropylacrylamide) (PNiPAAm) with its superior switching characteristic was paired with a poly(vinyl methyl ether)-based composition that allows adjusting physico-chemical and biomolecular properties in a wide range. Beyond pure PNiPAAm, the proposed thermo-responsive coating provides thickness, stiffness and swelling behavior, as well as an apposite density of reactive sites for biomolecular functionalization, as effective tuning parameters to meet specific requirements of a particular cell type regarding initial adhesion and ease of detachment. To illustrate the strength of this approach, the novel cell culture carrier was applied to generate transplantable sheets of human corneal endothelial cells (HCEC). Sheets were grown, detached, and transferred onto planar targets. Cell morphology, viability and functionality were analyzed by immunocytochemistry and determination of transepithelial electrical resistance (TEER) before and after sheet detachment and transfer. HCEC layers showed regular morphology with appropriate TEER. Cells were positive for function-associated marker proteins ZO-1, Na+/K+-ATPase, and paxillin, and extracellular matrix proteins fibronectin, laminin and collagen type IV before and after transfer. Sheet detachment and transfer did not impair cell viability. Subsequently, a potential application in ophthalmology was demonstrated by transplantation onto de-endothelialized porcine corneas in vitro. The novel thermo-responsive cell culture carrier facilitates the generation and transfer of functional HCEC sheets. This paves the way to generate tissue engineered human corneal endothelium as an alternative transplant source for endothelial keratoplasty.

Thermo-responsive cell culture carriers based on poly(vinyl methyl ether)-the effect of biomolecular ligands to balance cell adhesion and stimulated detachment.

PubMed

Teichmann, Juliane; Nitschke, Mirko; Pette, Dagmar; Valtink, Monika; Gramm, Stefan; Härtel, Frauke V; Noll, Thomas; Funk, Richard H W; Engelmann, Katrin; Werner, Carsten

2015-08-01

Two established material systems for thermally stimulated detachment of adherent cells were combined in a cross-linked polymer blend to merge favorable properties. Through this approach poly( N -isopropylacrylamide) (PNiPAAm) with its superior switching characteristic was paired with a poly(vinyl methyl ether)-based composition that allows adjusting physico-chemical and biomolecular properties in a wide range. Beyond pure PNiPAAm, the proposed thermo-responsive coating provides thickness, stiffness and swelling behavior, as well as an apposite density of reactive sites for biomolecular functionalization, as effective tuning parameters to meet specific requirements of a particular cell type regarding initial adhesion and ease of detachment. To illustrate the strength of this approach, the novel cell culture carrier was applied to generate transplantable sheets of human corneal endothelial cells (HCEC). Sheets were grown, detached, and transferred onto planar targets. Cell morphology, viability and functionality were analyzed by immunocytochemistry and determination of transepithelial electrical resistance (TEER) before and after sheet detachment and transfer. HCEC layers showed regular morphology with appropriate TEER. Cells were positive for function-associated marker proteins ZO-1, Na + /K + -ATPase, and paxillin, and extracellular matrix proteins fibronectin, laminin and collagen type IV before and after transfer. Sheet detachment and transfer did not impair cell viability. Subsequently, a potential application in ophthalmology was demonstrated by transplantation onto de-endothelialized porcine corneas in vitro . The novel thermo-responsive cell culture carrier facilitates the generation and transfer of functional HCEC sheets. This paves the way to generate tissue engineered human corneal endothelium as an alternative transplant source for endothelial keratoplasty.

Thermo-responsive cell culture carriers based on poly(vinyl methyl ether)—the effect of biomolecular ligands to balance cell adhesion and stimulated detachment

PubMed Central

Teichmann, Juliane; Nitschke, Mirko; Pette, Dagmar; Valtink, Monika; Gramm, Stefan; Härtel, Frauke V; Noll, Thomas; Funk, Richard H W; Engelmann, Katrin; Werner, Carsten

2015-01-01

Two established material systems for thermally stimulated detachment of adherent cells were combined in a cross-linked polymer blend to merge favorable properties. Through this approach poly(N-isopropylacrylamide) (PNiPAAm) with its superior switching characteristic was paired with a poly(vinyl methyl ether)-based composition that allows adjusting physico-chemical and biomolecular properties in a wide range. Beyond pure PNiPAAm, the proposed thermo-responsive coating provides thickness, stiffness and swelling behavior, as well as an apposite density of reactive sites for biomolecular functionalization, as effective tuning parameters to meet specific requirements of a particular cell type regarding initial adhesion and ease of detachment. To illustrate the strength of this approach, the novel cell culture carrier was applied to generate transplantable sheets of human corneal endothelial cells (HCEC). Sheets were grown, detached, and transferred onto planar targets. Cell morphology, viability and functionality were analyzed by immunocytochemistry and determination of transepithelial electrical resistance (TEER) before and after sheet detachment and transfer. HCEC layers showed regular morphology with appropriate TEER. Cells were positive for function-associated marker proteins ZO-1, Na+/K+-ATPase, and paxillin, and extracellular matrix proteins fibronectin, laminin and collagen type IV before and after transfer. Sheet detachment and transfer did not impair cell viability. Subsequently, a potential application in ophthalmology was demonstrated by transplantation onto de-endothelialized porcine corneas in vitro. The novel thermo-responsive cell culture carrier facilitates the generation and transfer of functional HCEC sheets. This paves the way to generate tissue engineered human corneal endothelium as an alternative transplant source for endothelial keratoplasty. PMID:27877823

Edaravone, an ROS Scavenger, Ameliorates Photoreceptor Cell Death after Experimental Retinal Detachment

PubMed Central

Roh, Mi In; Murakami, Yusuke; Thanos, Aristomenis; Miller, Joan W.

2011-01-01

Purpose. To investigate whether edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one), a free radical scavenger, would be neuroprotective against photoreceptor cell death in a rat model of retinal detachment (RD). Methods. RD was induced in adult Brown Norway rats by subretinal injection of sodium hyaluronate. Edaravone (3, 5, or 10 mg/kg) or physiologic saline was administered intraperitoneally once a day until death on day 3 or 5. Oxidative stress in the retina was assessed by 4-hydroxynonenal staining or ELISA for protein carbonyl content. Photoreceptor death was assessed by TUNEL and measurement of the outer nuclear layer thickness. Western blot analysis and caspase activity assays were performed. Inflammatory cytokine secretion and inflammatory cell infiltration were evaluated by ELISA and immunostaining, respectively. Results. RD resulted in increased generation of ROS. Treatment with 5 mg/kg edaravone significantly reduced the ROS level, along with a decrease in TUNEL-positive cells in the photoreceptor layer. A caspase assay also confirmed decreased activation of caspase-3, -8, and -9 in RD treated with edaravone. The level of the antiapoptotic Bcl-2 was increased in detached retinas after edaravone treatment, whereas the levels of the stress-activated p-ERK1/2 were decreased. In addition, edaravone treatment resulted in a significant decrease in the levels of TNF-α, MCP-1, and macrophage infiltration. Conclusions. Oxidative stress plays an important role in photoreceptor cell death after RD. Edaravone treatment may aid in preventing photoreceptor cell death after RD by suppressing ROS-induced photoreceptor damage. PMID:21310909

Detachment of affinity-captured bioparticles by elastic deformation of a macroporous hydrogel

PubMed Central

Dainiak, Maria B.; Kumar, Ashok; Galaev, Igor Yu.; Mattiasson, Bo

2006-01-01

Adsorption of bioparticles to affinity surfaces involves polyvalent interactions, complicating greatly the recovery of the adsorbed material. A unique system for the efficient binding and release of different cells and particles is described. Affinity-bound bioparticles and synthetic particles are detached from the macroporous hydrogel matrix, a so-called cryogel, when the cryogel undergoes elastic deformation. The particle detachment upon elastic deformation is believed to be due to breaking of many of the multipoint attachments between the particles and the affinity matrix and the change in the distance between affinity ligands when the matrix is deformed. However, no release of affinity-bound protein occurred upon elastic deformation. The phenomenon of particle detachment upon elastic deformation is believed to be of a generic nature, because it was demonstrated for a variety of bioparticles of different sizes and for synthetic particles, for different ligand–receptor pairs (IgG–protein A, sugar–ConA, metal ion–chelating ligand), and when the deformation was caused by either external forces (mechanical deformation) or internal forces (the shrinkage of thermosensitive, macroporous hydrogel upon an increase in temperature). The elasticity of cryogel monoliths ensures high recovery of captured cells under mild conditions, with highly retained viability. This property, along with their continuous porous structure makes cryogel monoliths very attractive for applications in affinity cell separation. PMID:16418282

Vitreous Microparticle Shedding in Retinal Detachment: A Prospective Comparative Study.

PubMed

Tumahai, Perle; Saas, Philippe; Ricouard, Fanny; Biichlé, Sabéha; Puyraveau, Marc; Laheurte, Caroline; Delbosc, Bernard; Saleh, Maher

2016-01-01

Microparticles (MPs) are membrane-derived vesicles measuring less than 1 μm in diameter. They are shed from nearly every activated or preapoptotic cell and may exhibit biologic activities in inflammation or apoptosis settings. The main purpose of this study was to determine whether MP shedding was higher in the vitreous of patients with retinal detachment (RD). This was a prospective, comparative study. Levels of vitreous MPs (including phosphatidylserine [PS]-expressing MPs, photoreceptor cell-derived MPs, and photoreceptor cell-derived MPs expressing PS) and soluble proinflammatory factors (i.e., monocyte chemoattractant protein-1, intercellular adhesion molecule-1, and IL-6) were analyzed by flow cytometry. Samples were obtained from 49 eyes undergoing RD surgery and 41 control eyes. Vitreous levels of all the MPs studied were significantly increased in the RD group. Vitreous MP levels were correlated with levels of at least one proinflammatory factor depending on MP subsets. Concerning clinical parameters, vitreous PS-expressing MP and PS-expressing photoreceptor cell-derived MP levels were higher depending on the duration of RD at surgery, the detached retina surface, and the macula status and were found more sensitive than proinflammatory factors only for the duration of RD at surgery. Vitreous concentrations of MPs (mainly derived from photoreceptor cells) are higher after rhegmatogenous RD and found to be correlated with soluble proinflammatory factors.

Inhibition of Ebola virus glycoprotein-mediated cytotoxicity by targeting its transmembrane domain and cholesterol.

PubMed

Hacke, Moritz; Björkholm, Patrik; Hellwig, Andrea; Himmels, Patricia; Ruiz de Almodóvar, Carmen; Brügger, Britta; Wieland, Felix; Ernst, Andreas M

2015-07-09

The high pathogenicity of the Ebola virus reflects multiple concurrent processes on infection. Among other important determinants, Ebola fusogenic glycoprotein (GP) has been associated with the detachment of infected cells and eventually leads to vascular leakage and haemorrhagic fever. Here we report that the membrane-anchored GP is sufficient to induce the detachment of adherent cells. The results show that the detachment induced through either full-length GP1,2 or the subunit GP2 depends on cholesterol and the structure of the transmembrane domain. These data reveal a novel molecular mechanism in which GP regulates Ebola virus assembly and suggest that cholesterol-reducing agents could be useful as therapeutics to counteract GP-mediated cell detachment.

Separation of abscission zone cells in detached Azolla roots depends on apoplastic pH.

PubMed

Fukuda, Kazuma; Yamada, Yoshiya; Miyamoto, Kensuke; Ueda, Junichi; Uheda, Eiji

2013-01-01

In studies on the mechanism of cell separation during abscission, little attention has been paid to the apoplastic environment. We found that the apoplastic pH surrounding abscission zone cells in detached roots of the water fern Azolla plays a major role in cell separation. Abscission zone cells of detached Azolla roots were separated rapidly in a buffer at neutral pH and slowly in a buffer at pH below 4.0. However, cell separation rarely occurred at pH 5.0-5.5. Light and electron microscopy revealed that cell separation was caused by a degradation of the middle lamella between abscission zone cells at both pH values, neutral and below 4.0. Low temperature and papain treatment inhibited cell separation. Enzyme(s) in the cell wall of the abscission zone cells might be involved in the degradation of the pectin of the middle lamella and the resultant, pH-dependent cell separation. By contrast, in Phaseolus leaf petioles, unlike Azolla roots, cell separation was slow and increased only at acidic pH. The rapid cell separation, as observed in Azolla roots at neutral pH, did not occur. Indirect immunofluorescence microscopy, using anti-pectin monoclonal antibodies, revealed that the cell wall pectins of the abscission zone cells of Azolla roots and Phaseolus leaf petioles looked similar and changed similarly during cell separation. Thus, the pH-related differences in cell separation mechanisms of Azolla and Phaseolus might not be due to differences in cell wall pectin, but to differences in cell wall-located enzymatic activities responsible for the degradation of pectic substances. A possible enzyme system is discussed. Copyright © 2012 Elsevier GmbH. All rights reserved.

Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment.

PubMed

Kobayashi, Jun; Hayashi, Masaki; Ohno, Takahiro; Nishi, Masanori; Arisaka, Yoshinori; Matsubara, Yoshinori; Kakidachi, Hiroshi; Akiyama, Yoshikatsu; Yamato, Masayuki; Horii, Akihiro; Okano, Teruo

2014-11-01

Antibody-immobilized thermoresponsive poly(N-isopropylacrylamide-co-2-carboxyisopropylacrylamide) [poly(IPAAm-co-CIPAAm)]-grafted cell culture surfaces were designed to enhance both the initial adhesion of weakly adhering cells and the ability of cells to detach in response to low temperature through the regulation of affinity binding between immobilized antibodies and antigens on the cellular surface. Ty-82 cells and neonatal normal human dermal fibroblasts (NHDFs), which express CD90 on the cell surface, adhered to anti-CD90 antibody-immobilized thermoresponsive surfaces at 37°C, a condition at which the grafted thermoresponsive polymer chains shrank. Adherent Ty-82 cells were detached from the surfaces by lowering the temperature to 20°C and applying external forces, such as pipetting, whereas cultured NHDF sheets spontaneously detached themselves from the surface in response to reduced temperature alone. When the temperature was decreased to 20°C, the swelling of grafted thermoresponsive polymer chains weakened the affinity binding between immobilized antibody and antigen on the cells due to the increasing steric hindrance of the polymer chains around the antigen-recognition site of the immobilized antibodies. No contamination was detected on cells harvested from covalently immobilized antibodies on the culture surfaces by low-temperature treatment, whereas a carryover of the antibody and avidin from the avidin-biotin binding surface was observed. Furthermore, the initial adhesion of adipose tissue-derived cells, which adhere weakly to PIPAAm-grafted surfaces, was enhanced on the antibody-immobilized thermoresponsive surfaces. © 2013 Wiley Periodicals, Inc.

A New In Vitro Model of Breast Cancer Metastasis to Bone

DTIC Science & Technology

2010-04-01

excessive cell vacuolization, ruffled cell margins , and increasing rates of cell-surface detachment. Thus, conventional tissue-culture strategies...determined. 3050 DHURJATI ET AL. apoptotic bodies, cytoplasmic vesiculation, and chromatin margination (Fig. 3) and a distinct loss of insoluble ECM...respond abnormally to various hor- mones and cytokines compared to normal, differen- tiated human osteoblasts. In effort to overcome these limitations

Calcium-Oxidant Signaling Network Regulates AMP-activated Protein Kinase (AMPK) Activation upon Matrix Deprivation*

PubMed Central

Sundararaman, Ananthalakshmy; Amirtham, Usha; Rangarajan, Annapoorni

2016-01-01

The AMP-activated protein kinase (AMPK) has recently been implicated in anoikis resistance. However, the molecular mechanisms that activate AMPK upon matrix detachment remain unexplored. In this study, we show that AMPK activation is a rapid and sustained phenomenon upon matrix deprivation, whereas re-attachment to the matrix leads to its dephosphorylation and inactivation. Because matrix detachment leads to loss of integrin signaling, we investigated whether integrin signaling negatively regulates AMPK activation. However, modulation of focal adhesion kinase or Src, the major downstream components of integrin signaling, failed to cause a corresponding change in AMPK signaling. Further investigations revealed that the upstream AMPK kinases liver kinase B1 (LKB1) and Ca2+/calmodulin-dependent protein kinase kinase β (CaMKKβ) contribute to AMPK activation upon detachment. In LKB1-deficient cells, we found AMPK activation to be predominantly dependent on CaMKKβ. We observed no change in ATP levels under detached conditions at early time points suggesting that rapid AMPK activation upon detachment was not triggered by energy stress. We demonstrate that matrix deprivation leads to a spike in intracellular calcium as well as oxidant signaling, and both these intracellular messengers contribute to rapid AMPK activation upon detachment. We further show that endoplasmic reticulum calcium release-induced store-operated calcium entry contributes to intracellular calcium increase, leading to reactive oxygen species production, and AMPK activation. We additionally show that the LKB1/CaMKK-AMPK axis and intracellular calcium levels play a critical role in anchorage-independent cancer sphere formation. Thus, the Ca2+/reactive oxygen species-triggered LKB1/CaMKK-AMPK signaling cascade may provide a quick, adaptable switch to promote survival of metastasizing cancer cells. PMID:27226623

The Globoside Receptor Triggers Structural Changes in the B19 Virus Capsid That Facilitate Virus Internalization▿

PubMed Central

Bönsch, Claudia; Zuercher, Christoph; Lieby, Patricia; Kempf, Christoph; Ros, Carlos

2010-01-01

Globoside (Gb4Cer), Ku80 autoantigen, and α5β1 integrin have been identified as cell receptors/coreceptors for human parvovirus B19 (B19V), but their role and mechanism of interaction with the virus are largely unknown. In UT7/Epo cells, expression of Gb4Cer and CD49e (integrin alpha-5) was high, but expression of Ku80 was insignificant. B19V colocalized with Gb4Cer and, to a lesser extent, with CD49e. However, only anti-Gb4Cer antibodies could disturb virus attachment. Only a small proportion of cell-bound viruses were internalized, while the majority became detached from the receptor. When added to uninfected cells, the receptor-detached virus showed superior cell binding capacity and infectivity. Attachment of B19V to cells triggered conformational changes in the capsid leading to the accessibility of the N terminus of VP1 (VP1u) to antibodies, which was maintained in the receptor-detached virus. VP1u became similarly accessible to antibodies following incubation of B19V particles with increasing concentrations of purified Gb4Cer. The receptor-mediated exposure of VP1u is critical for virus internalization, since capsids lacking VP1 could bind to cells but were not internalized. Moreover, an antibody against the N terminus of VP1u disturbed virus internalization, but only when present during and not after virus attachment, indicating the involvement of this region in binding events required for internalization. These results suggest that Gb4Cer is not only the primary receptor for B19V attachment but also the mediator of capsid rearrangements required for subsequent interactions leading to virus internalization. The capacity of the virus to detach and reattach again would enhance the probability of productive infections. PMID:20826697

Multicellular detachment generates metastatic spheroids during intra-abdominal dissemination in epithelial ovarian cancer.

PubMed

Al Habyan, Sara; Kalos, Christina; Szymborski, Joseph; McCaffrey, Luke

2018-05-23

Ovarian cancer is the most lethal gynecological cancer, where survival rates have had modest improvement over the last 30 years. Metastasis of cancer cells is a major clinical problem, and patient mortality occurs when ovarian cancer cells spread beyond the confinement of ovaries. Disseminated ovarian cancer cells typically spread within the abdomen, where ascites accumulation aids in their transit. Metastatic ascites contain multicellular spheroids, which promote chemo-resistance and recurrence. However, little is known about the origin and mechanisms through which spheroids arise. Using live-imaging of 3D culture models and animal models, we report that epithelial ovarian cancer (EOC) cells, the most common type of ovarian cancer, can spontaneously detach as either single cells or clusters. We report that clusters are more resistant to anoikis and have a potent survival advantage over single cells. Using in vivo lineage tracing, we found that multicellular spheroids arise preferentially from collective detachment, rather than aggregation in the abdomen. Finally, we report that multicellular spheroids from collective detachment are capable of seeding intra-abdominal metastases that retain intra-tumoral heterogeneity from the primary tumor.

Exogenous ROS-induced cell sheet transfer based on hematoporphyrin-polyketone film via a one-step process.

PubMed

Koo, Min-Ah; Lee, Mi Hee; Kwon, Byeong-Ju; Seon, Gyeung Mi; Kim, Min Sung; Kim, Dohyun; Nam, Ki Chang; Park, Jong-Chul

2018-04-01

To date, most of invasive cell sheet harvesting methods have used culture surface property variations, such as wettability, pH, electricity, and magnetism, to induce cell detachment. These methods that rely on surface property changes are effective when cell detachment prior to application is necessary, but of limited use when used for cell sheet transfer to target regions. The study reports a new reactive oxygen species (ROS)-induced strategy based on hematoporphyrin-incorporated polyketone film (Hp-PK film) to transfer cell sheets directly to target areas without an intermediate harvesting process. After green LED (510 nm) irradiation, production of exogenous ROS from the Hp-PK films induces cell sheet detachment and transfer. The study suggests that ROS-induced cell detachment property of the Hp-PK film is closely related to conformational changes of extracellular matrix (ECM) proteins. Also, this strategy with the Hp-PK film can be applied by regulating production rate of exogenous ROS in various types of cells, including fibroblasts, mesenchymal stem cells and keratinocytes. In conclusion, ROS-induced method using the Hp-PK film can be used for one-step cell sheet transplantation and has potential in biomedical applications. Copyright © 2018 Elsevier Ltd. All rights reserved.

Proliferative vitreoretinopathy in the Swine-a new model.

PubMed

Umazume, Kazuhiko; Barak, Yoreh; McDonald, Kevin; Liu, Lanhsin; Kaplan, Henry J; Tamiya, Shigeo

2012-07-24

To develop a large animal model of proliferative vitreoretinopathy (PVR) in the swine to eventually study disease pathophysiology, as well as novel therapies. PVR was induced in domestic swine by creation of a posterior vitreous detachment, creation of a retinal detachment by the injection of subretinal fluid, and intravitreal injection of green fluorescent protein-positive retinal pigment epithelial (GFP+ RPE) cells. Control eyes had the same surgical procedures without RPE cell injection. PVR was clinically graded on days 3, 7, and 14. Animals were euthanized on day 14, and enucleated eyes were analyzed by light microscopy and immunohistochemistry. Injection of GFP+ RPE cells into the vitreous cavity produced localized, traction retinal detachments by day 14 in all eyes (14 of 14); in contrast, the retina spontaneously reattached by day 3 and remained attached in all control eyes (10 of 10). Contractile epiretinal membranes on the inner retinal surface that caused the traction retinal detachments consisted predominantly of GFP+ RPE cells. These cells stained positive for cytokeratin, confirming their epithelial origin, and also expressed α-SMA and fibronectin, markers for myofibroblasts and fibrosis, respectively. We established a swine PVR model that recapitulates key clinical features found in humans and, thus, can be used to study the pathophysiology of PVR, as well as new novel therapies. GFP+ RPE cells injected into the vitreous cavity formed contractile membranes on the inner retinal surface and caused localized traction retinal detachments.

Affinity adsorption of cells to surfaces and strategies for cell detachment.

PubMed

Hubble, John

2007-01-01

The use of bio-specific interactions for the separation and recovery of bio-molecules is now widely established and in many cases the technique has successfully crossed the divide between bench and process scale operation. Although the major specificity advantage of affinity-based separations also applies to systems intended for cell fractionation, developments in this area have been slower. Many of the problems encountered result from attempts to take techniques developed for molecular systems and, with only minor modification to the conditions used, apply them for the separation of cells. This approach tends to ignore or at least trivialise the problems, which arise from the heterogeneous nature of a cell suspension and the multivalent nature of the cell/surface interaction. To develop viable separation processes on a larger scale, effective contacting strategies are required in separators that also allow detachment or recovery protocols that overcome the enhanced binding strength generated by multivalent interactions. The effects of interaction valency on interaction strength needs to be assessed and approaches developed to allow effective detachment and recovery of adsorbed cells without compromising cell viability. This article considers the influence of operating conditions on cell attachment and the extent to which multivalent interactions determine the strength of cell binding and subsequent detachment.

Biofilms’ Role in Planktonic Cell Proliferation

PubMed Central

Bester, Elanna; Wolfaardt, Gideon M.; Aznaveh, Nahid B.; Greener, Jesse

2013-01-01

The detachment of single cells from biofilms is an intrinsic part of this surface-associated mode of bacterial existence. Pseudomonas sp. strain CT07gfp biofilms, cultivated in microfluidic channels under continuous flow conditions, were subjected to a range of liquid shear stresses (9.42 mPa to 320 mPa). The number of detached planktonic cells was quantified from the effluent at 24-h intervals, while average biofilm thickness and biofilm surface area were determined by confocal laser scanning microscopy and image analysis. Biofilm accumulation proceeded at the highest applied shear stress, while similar rates of planktonic cell detachment was maintained for biofilms of the same age subjected to the range of average shear rates. The conventional view of liquid-mediated shear leading to the passive erosion of single cells from the biofilm surface, disregards the active contribution of attached cell metabolism and growth to the observed detachment rates. As a complement to the conventional conceptual biofilm models, the existence of a biofilm surface-associated zone of planktonic cell proliferation is proposed to highlight the need to expand the traditional perception of biofilms as promoting microbial survival, to include the potential of biofilms to contribute to microbial proliferation. PMID:24201127

Revealing Early Steps of α2β1 Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

PubMed Central

Taubenberger, Anna; Cisneros, David A.; Friedrichs, Jens; Puech, Pierre-Henri; Muller, Daniel J.

2007-01-01

We have characterized early steps of α2β1 integrin-mediated cell adhesion to a collagen type I matrix by using single-cell force spectroscopy. In agreement with the role of α2β1 as a collagen type I receptor, α2β1-expressing Chinese hamster ovary (CHO)-A2 cells spread rapidly on the matrix, whereas α2β1-negative CHO wild-type cells adhered poorly. Probing CHO-A2 cell detachment forces over a contact time range of 600 s revealed a nonlinear adhesion response. During the first 60 s, cell adhesion increased slowly, and forces associated with the smallest rupture events were consistent with the breakage of individual integrin–collagen bonds. Above 60 s, a fraction of cells rapidly switched into an activated adhesion state marked by up to 10-fold increased detachment forces. Elevated overall cell adhesion coincided with a rise of the smallest rupture forces above the value required to break a single-integrin–collagen bond, suggesting a change from single to cooperative receptor binding. Transition into the activated adhesion mode and the increase of the smallest rupture forces were both blocked by inhibitors of actomyosin contractility. We therefore propose a two-step mechanism for the establishment of α2β1-mediated adhesion as weak initial, single-integrin–mediated binding events are superseded by strong adhesive interactions involving receptor cooperativity and actomyosin contractility. PMID:17314408

Astrocyte structural reactivity and plasticity in models of retinal detachment.

PubMed

Luna, Gabriel; Keeley, Patrick W; Reese, Benjamin E; Linberg, Kenneth A; Lewis, Geoffrey P; Fisher, Steven K

2016-09-01

Although retinal neurodegenerative conditions such as age-related macular degeneration, glaucoma, diabetic retinopathy, retinitis pigmentosa, and retinal detachment have different etiologies and pathological characteristics, they also have many responses in common at the cellular level, including neural and glial remodeling. Structural changes in Müller cells, the large radial glia of the retina in retinal disease and injury have been well described, that of the retinal astrocytes remains less so. Using modern imaging technology to describe the structural remodeling of retinal astrocytes after retinal detachment is the focus of this paper. We present both a review of critical literature as well as novel work focusing on the responses of astrocytes following rhegmatogenous and serous retinal detachment. The mouse presents a convenient model system in which to study astrocyte reactivity since the Mϋller cell response is muted in comparison to other species thereby allowing better visualization of the astrocytes. We also show data from rat, cat, squirrel, and human retina demonstrating similarities and differences across species. Our data from immunolabeling and dye-filling experiments demonstrate previously undescribed morphological characteristics of normal astrocytes and changes induced by detachment. Astrocytes not only upregulate GFAP, but structurally remodel, becoming increasingly irregular in appearance, and often penetrating deep into neural retina. Understanding these responses, their consequences, and what drives them may prove to be an important component in improving visual outcome in a variety of therapeutic situations. Our data further supports the concept that astrocytes are important players in the retina's overall response to injury and disease. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Bud detachment in hydra requires activation of fibroblast growth factor receptor and a Rho–ROCK–myosin II signaling pathway to ensure formation of a basal constriction

PubMed Central

Holz, Oliver; Apel, David; Steinmetz, Patrick; Lange, Ellen; Hopfenmüller, Simon; Ohler, Kerstin; Sudhop, Stefanie

2017-01-01

Background: Hydra propagates asexually by exporting tissue into a bud, which detaches 4 days later as a fully differentiated young polyp. Prerequisite for detachment is activation of fibroblast growth factor receptor (FGFR) signaling. The mechanism which enables constriction and tissue separation within the monolayered ecto‐ and endodermal epithelia is unknown. Results: Histological sections and staining of F‐actin by phalloidin revealed conspicuous cell shape changes at the bud detachment site indicating a localized generation of mechanical forces and the potential enhancement of secretory functions in ectodermal cells. By gene expression analysis and pharmacological inhibition, we identified a candidate signaling pathway through Rho, ROCK, and myosin II, which controls bud base constriction and rearrangement of the actin cytoskeleton. Specific regional myosin phosphorylation suggests a crucial role of ectodermal cells at the detachment site. Inhibition of FGFR, Rho, ROCK, or myosin II kinase activity is permissive for budding, but represses myosin phosphorylation, rearrangement of F‐actin and constriction. The young polyp remains permanently connected to the parent by a broad tissue bridge. Conclusions: Our data suggest an essential role of FGFR and a Rho‐ROCK‐myosin II pathway in the control of cell shape changes required for bud detachment. Developmental Dynamics 246:502–516, 2017. © 2017 The Authors Developmental Dynamics published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists PMID:28411398

Bud detachment in hydra requires activation of fibroblast growth factor receptor and a Rho-ROCK-myosin II signaling pathway to ensure formation of a basal constriction.

PubMed

Holz, Oliver; Apel, David; Steinmetz, Patrick; Lange, Ellen; Hopfenmüller, Simon; Ohler, Kerstin; Sudhop, Stefanie; Hassel, Monika

2017-07-01

Hydra propagates asexually by exporting tissue into a bud, which detaches 4 days later as a fully differentiated young polyp. Prerequisite for detachment is activation of fibroblast growth factor receptor (FGFR) signaling. The mechanism which enables constriction and tissue separation within the monolayered ecto- and endodermal epithelia is unknown. Histological sections and staining of F-actin by phalloidin revealed conspicuous cell shape changes at the bud detachment site indicating a localized generation of mechanical forces and the potential enhancement of secretory functions in ectodermal cells. By gene expression analysis and pharmacological inhibition, we identified a candidate signaling pathway through Rho, ROCK, and myosin II, which controls bud base constriction and rearrangement of the actin cytoskeleton. Specific regional myosin phosphorylation suggests a crucial role of ectodermal cells at the detachment site. Inhibition of FGFR, Rho, ROCK, or myosin II kinase activity is permissive for budding, but represses myosin phosphorylation, rearrangement of F-actin and constriction. The young polyp remains permanently connected to the parent by a broad tissue bridge. Our data suggest an essential role of FGFR and a Rho-ROCK-myosin II pathway in the control of cell shape changes required for bud detachment. Developmental Dynamics 246:502-516, 2017. © 2017 The Authors Developmental Dynamics published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists. © 2017 The Authors Developmental Dynamics published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists.

Synergistic actions of pemphigus vulgaris IgG, Fas-ligand and tumor necrosis factor-alpha during induction of basal cell shrinkage and acantholysis.

PubMed

Orlov, Maxim D; Chernyavsky, Alex I; Arredondo, Juan; Grando, Sergei A

2006-11-01

This study tested a recently proposed "Basal Cell Shrinkage" hypothesis of pemphigus acantholysis through a quantitative analysis of individual and cooperative effects of pemphigus vulgaris (PV) IgG, Fas-ligand (Fas-L) and tumor necrosis factor-alpha (TNFalpha) on keratinocyte (KC) volume (i.e. cell size) and adhesive properties. Exposure of KC monolayers and MatTek EpiDermFT tissues cultures to the physiologic concentrations of Fas-L, TNFalpha or IgGs from two PV patients resulted in various degrees of reversible changes, which were not observed in control cultures either exposed to normal IgG or left intact. Within 12-24 h of exposure, basal cells in experimental cultures lost their ability to form stress fibers, retracted cytoplasmic aprons and formed keratin aggregates, indicating that their cytoskeleton collapsed. The cell volume decreased significantly (p < 0.05) as the polygonal cell shape changed to a round one. The shrunk cells detached from their neighbors and the substrate, resulting in a reciprocal increase of both the areas of acantholysis and the number of detached KCs, respectively. Since in the skin of PV patients, KCs are targeted by autoantibodies concomitantly with being exposed to autocrine and paracrine pro-apoptotic and pro-inflammatory cytokines, we combined PV IgG with Fas-L and/or TNFalpha in the cell culture experiments. This amplified several fold an ability of PV IgG to cause basal cell shrinkage and detachment. The obtained results demonstrated for the first time that PV IgG works together with Fas-L and TNFalpha to induce acantholysis via basal cell shrinkage, which provides a novel mechanism explaining successful treatment of PV patients with TNFalpha inhibitors.

Fundus autofluorescence findings in a mouse model of retinal detachment.

PubMed

Secondi, Roberta; Kong, Jian; Blonska, Anna M; Staurenghi, Giovanni; Sparrow, Janet R

2012-08-07

Fundus autofluorescence (fundus AF) changes were monitored in a mouse model of retinal detachment (RD). RD was induced by transscleral injection of hyaluronic acid (Healon) or sterile balanced salt solution (BSS) into the subretinal space of 4-5-day-old albino Abca4 null mutant and Abca4 wild-type mice. Images acquired by confocal scanning laser ophthalmoscopy (Spectralis HRA) were correlated with spectral domain optical coherence tomography (SD-OCT), infrared reflectance (IR), fluorescence spectroscopy, and histologic analysis. Results. In the area of detached retina, multiple hyperreflective spots in IR images corresponded to punctate areas of intense autofluorescence visible in fundus AF mode. The puncta exhibited changes in fluorescence intensity with time. SD-OCT disclosed undulations of the neural retina and hyperreflectivity of the photoreceptor layer that likely corresponded to histologically visible photoreceptor cell rosettes. Fluorescence emission spectra generated using flat-mounted retina, and 488 and 561 nm excitation, were similar to that of RPE lipofuscin. With increased excitation wavelength, the emission maximum shifted towards longer wavelengths, a characteristic typical of fundus autofluorescence. In detached retinas, hyper-autofluorescent spots appeared to originate from photoreceptor outer segments that were arranged within retinal folds and rosettes. Consistent with this interpretation is the finding that the autofluorescence was spectroscopically similar to the bisretinoids that constitute RPE lipofuscin. Under the conditions of a RD, abnormal autofluorescence may arise from excessive production of bisretinoid by impaired photoreceptor cells.

Mangiferin protects mitochondrial function by preserving mitochondrial hexokinase-II in vessel endothelial cells.

PubMed

Song, Junna; Li, Yi; Song, Junmei; Hou, Fangjie; Liu, Baolin; Li, Aiying

2017-07-01

Hexokinase-II (HK-II) confers protection against cell death and this study was designed to investigate the effect of mangiferin on the regulation of mitochondrial HK-II. In vessel endothelial cells, saturated fatty acid palmitate (PA) stimulation induced HK-II detachment from mitochondria due to cellular acidification. Mangiferin reduced lactate accumulation by improving pyruvate dehydrogenase activity, promoted Akt translocation to HK-II and prevented HK-II detachment from mitochondria. Knockdown of Akt2 diminished the protective effect of mangiferin on mitochondrial HK-II, confirming the role of Akt in the regulation of HK-II. Mangiferin prevented mitochondrial permeability transition pore opening, restored mitochondrial membrane potential and thereby protected cell from apoptosis. In high-fat diet fed mice, oral administration of mangiferin induced Akt phosphorylation, increased HK-II binding to mitochondria and resultantly protected vessel endothelial function, demonstrating its protective effect on endothelial integrity in vivo. This finding provided a novel strategy for the protection of mitochondrial function in the endothelium. Copyright © 2017 Elsevier B.V. All rights reserved.

Taurolithocholate-induced MRP2 retrieval involves MARCKS phosphorylation by protein kinase Cϵ in HUH-NTCP Cells.

PubMed

Schonhoff, Christopher M; Webster, Cynthia R L; Anwer, M Sawkat

2013-07-01

Taurolithocholate (TLC) acutely inhibits the biliary excretion of multidrug-resistant associated protein 2 (Mrp2) substrates by inducing Mrp2 retrieval from the canalicular membrane, whereas cyclic adenosine monophosphate (cAMP) increases plasma membrane (PM)-MRP2. The effect of TLC may be mediated via protein kinase Cϵ (PKCϵ). Myristoylated alanine-rich C kinase substrate (MARCKS) is a membrane-bound F-actin crosslinking protein and is phosphorylated by PKCs. MARCKS phosphorylation has been implicated in endocytosis, and the underlying mechanism appears to be the detachment of phosphorylated myristoylated alanine-rich C kinase substrate (pMARCKS) from the membrane. The aim of the present study was to test the hypothesis that TLC-induced MRP2 retrieval involves PKCϵ-mediated MARCKS phosphorylation. Studies were conducted in HuH7 cells stably transfected with sodium taurocholate cotransporting polypeptide (HuH-NTCP cells) and in rat hepatocytes. TLC increased PM-PKCϵ and decreased PM-MRP2 in both HuH-NTCP cells and hepatocytes. cAMP did not affect PM-PKCϵ and increased PM-MRP2 in these cells. In HuH-NTCP cells, dominant-negative (DN) PKCϵ reversed TLC-induced decreases in PM-MRP2 without affecting cAMP-induced increases in PM-MRP2. TLC, but not cAMP, increased MARCKS phosphorylation in HuH-NTCP cells and hepatocytes. TLC and phorbol myristate acetate increased cytosolic pMARCKS and decreased PM-MARCKS in HuH-NTCP cells. TLC failed to increase MARCKS phosphorylation in HuH-NTCP cells transfected with DN-PKCϵ, and this suggested PKCϵ-mediated phosphorylation of MARCKS by TLC. In HuH-NTCP cells transfected with phosphorylation-deficient MARCKS, TLC failed to increase MARCKS phosphorylation or decrease PM-MRP2. Taken together, these results support the hypothesis that TLC-induced MRP2 retrieval involves TLC-mediated activation of PKCϵ followed by MARCKS phosphorylation and consequent detachment of MARCKS from the membrane. Copyright © 2013 American Association for the Study of Liver Diseases.

Distinct molecular and cellular contributions to stabilizing selectin-mediated rolling under flow

PubMed Central

Yago, Tadayuki; Leppänen, Anne; Qiu, Haiying; Marcus, Warren D.; Nollert, Matthias U.; Zhu, Cheng; Cummings, Richard D.; McEver, Rodger P.

2002-01-01

Leukocytes roll on selectins at nearly constant velocities over a wide range of wall shear stresses. Ligand-coupled microspheres roll faster on selectins and detach quickly as wall shear stress is increased. To examine whether the superior performance of leukocytes reflects molecular features of native ligands or cellular properties that favor selectin-mediated rolling, we coupled structurally defined selectin ligands to microspheres or K562 cells and compared their rolling on P-selectin. Microspheres bearing soluble P-selectin glycoprotein ligand (sPSGL)-1 or 2-glycosulfopeptide (GSP)-6, a GSP modeled after the NH2-terminal P-selectin–binding region of PSGL-1, rolled equivalently but unstably on P-selectin. K562 cells displaying randomly coupled 2-GSP-6 also rolled unstably. In contrast, K562 cells bearing randomly coupled sPSGL-1 or 2-GSP-6 targeted to a membrane-distal region of the presumed glycocalyx rolled more like leukocytes: rolling steps were more uniform and shear resistant, and rolling velocities tended to plateau as wall shear stress was increased. K562 cells treated with paraformaldehyde or methyl-β-cyclodextrin before ligand coupling were less deformable and rolled unstably like microspheres. Cells treated with cytochalasin D were more deformable, further resisted detachment, and rolled slowly despite increases in wall shear stress. Thus, stable, shear-resistant rolling requires cellular properties that optimize selectin–ligand interactions. PMID:12177042

Effects of membrane deformability and bond formation/dissociation rates on adhesion dynamics of a spherical capsule in shear flow.

PubMed

Zhang, Ziying; Du, Jun; Wei, Zhengying; Wang, Zhen; Li, Minghui

2018-02-01

Cellular adhesion plays a critical role in biological systems and biomedical applications. Cell deformation and biophysical properties of adhesion molecules are of significance for the adhesion behavior. In the present work, dynamic adhesion of a deformable capsule to a planar substrate, in a linear shear flow, is numerically simulated to investigate the combined influence of membrane deformability (quantified by the capillary number) and bond formation/dissociation rates on the adhesion behavior. The computational model is based on the immersed boundary-lattice Boltzmann method for the capsule-fluid interaction and a probabilistic adhesion model for the capsule-substrate interaction. Three distinct adhesion states, detachment, rolling adhesion and firm adhesion, are identified and presented in a state diagram as a function of capillary number and bond dissociation rate. The impact of bond formation rate on the state diagram is further investigated. Results show that the critical bond dissociation rate for the transition of rolling or firm adhesion to detachment is strongly related to the capsule deformability. At the rolling-adhesion state, smaller off rates are needed for larger capillary number to increase the rolling velocity and detach the capsule. In contrast, the critical off rate for firm-to-detach transition slightly increases with the capillary number. With smaller on rate, the effect of capsule deformability on the critical off rates is more pronounced and capsules with moderate deformability are prone to detach by the shear flow. Further increasing of on rate leads to large expansion of both rolling-adhesion and firm-adhesion regions. Even capsules with relatively large deformability can maintain stable rolling adhesion at certain off rate.

Quantifying the effect of electric current on cell adhesion studied by single-cell force spectroscopy.

PubMed

Jaatinen, Leena; Young, Eleanore; Hyttinen, Jari; Vörös, János; Zambelli, Tomaso; Demkó, László

2016-03-20

This study presents the effect of external electric current on the cell adhesive and mechanical properties of the C2C12 mouse myoblast cell line. Changes in cell morphology, viability, cytoskeleton, and focal adhesion structure were studied by standard staining protocols, while single-cell force spectroscopy based on the fluidic force microscopy technology provided a rapid, serial quantification and detailed analysis of cell adhesion and its dynamics. The setup allowed measurements of adhesion forces up to the μN range, and total detachment distances over 40 μm. Force-distance curves have been fitted with a simple elastic model including a cell detachment protocol in order to estimate the Young's modulus of the cells, as well as to reveal changes in the dynamic properties as functions of the applied current dose. While the cell spreading area decreased monotonously with increasing current doses, small current doses resulted only in differences related to cell elasticity. Current doses above 11 As/m(2), however, initiated more drastic changes in cell morphology, viability, cellular structure, as well as in properties related to cell adhesion. The observed differences, eventually leading to cell death toward higher doses, might originate from both the decrease in pH and the generation of reactive oxygen species.

Single-cell adhesion probed in-situ using optical tweezers: A case study with Saccharomyces cerevisiae

NASA Astrophysics Data System (ADS)

Castelain, Mickaël; Rouxhet, Paul G.; Pignon, Frédéric; Magnin, Albert; Piau, Jean-Michel

2012-06-01

A facile method of using optical trapping to measure cell adhesion forces is presented and applied to the adhesion of Saccharomyces cerevisiae on glass, in contact with solutions of different compositions. Trapping yeast cells with optical tweezers (OT) is not perturbed by cell wall deformation or cell deviation from a spherical shape. The trapping force calibration requires correction not only for the hydrodynamic effect of the neighboring wall but also for spherical aberrations affecting the focal volume and the trap stiffness. Yeast cells trapped for up to 5 h were still able to undergo budding but showed an increase of doubling time. The proportion of adhering cells showed the expected variation according to the solution composition. The detachment force varied in the same way. This observation and the fact that the detachment stress was exerted parallel to the substrate surface point to the role of interactions involving solvated macromolecules. Both the proportion of adhering cells and the removal force showed a distribution which, in our experimental conditions, must be attributed to a heterogeneity of surface properties at the cell level or at the subcellular scale. As compared with magnetic tweezers, atomic force microscopy, and more conventional ways of studying cell adhesion (shear-flow cells), OT present several advantages that are emphasized in this paper.

Bilateral orbital infarction and retinal detachment in a previously undiagnosed sickle cell hemoglobinopathy African child

PubMed Central

Helen, Onakpoya Oluwatoyin; Ajite, K. O.; Oyelami, O. A.; Asaleye, C. M.; Adeoye, A. O.

2013-01-01

Bone infarction involving the orbit in sickle cell disease is not common. Bilateral orbital infarction in a previously undiagnosed sickle cell hemoglobinopathy has not been previously reported. In this report, we present a case of an 11-year-old previously undiagnosed sickle cell disease Nigerian girl with severe acute bilateral orbital infarction and retinal detachment to highlight that hemoglobinopathy induced orbital infarction should be considered in African children with acute onset proptosis with or without previous history of sickle cell hemoglobinopathy. PMID:23901183

Fundus Autofluorescence Findings in a Mouse Model of Retinal Detachment

PubMed Central

Secondi, Roberta; Kong, Jian; Blonska, Anna M.; Staurenghi, Giovanni; Sparrow, Janet R.

2012-01-01

Purpose. Fundus autofluorescence (fundus AF) changes were monitored in a mouse model of retinal detachment (RD). Methods. RD was induced by transscleral injection of hyaluronic acid (Healon) or sterile balanced salt solution (BSS) into the subretinal space of 4–5-day-old albino Abca4 null mutant and Abca4 wild-type mice. Images acquired by confocal scanning laser ophthalmoscopy (Spectralis HRA) were correlated with spectral domain optical coherence tomography (SD-OCT), infrared reflectance (IR), fluorescence spectroscopy, and histologic analysis. Results. In the area of detached retina, multiple hyperreflective spots in IR images corresponded to punctate areas of intense autofluorescence visible in fundus AF mode. The puncta exhibited changes in fluorescence intensity with time. SD-OCT disclosed undulations of the neural retina and hyperreflectivity of the photoreceptor layer that likely corresponded to histologically visible photoreceptor cell rosettes. Fluorescence emission spectra generated using flat-mounted retina, and 488 and 561 nm excitation, were similar to that of RPE lipofuscin. With increased excitation wavelength, the emission maximum shifted towards longer wavelengths, a characteristic typical of fundus autofluorescence. Conclusions. In detached retinas, hyper-autofluorescent spots appeared to originate from photoreceptor outer segments that were arranged within retinal folds and rosettes. Consistent with this interpretation is the finding that the autofluorescence was spectroscopically similar to the bisretinoids that constitute RPE lipofuscin. Under the conditions of a RD, abnormal autofluorescence may arise from excessive production of bisretinoid by impaired photoreceptor cells. PMID:22786896

Optimization of low energy sonication treatment for granular activated carbon colonizing biomass assessment.

PubMed

Saccani, G; Bernasconi, M; Antonelli, M

2014-01-01

This study is aimed at optimizing a low energy sonication (LES) treatment for granular activated carbon (GAC)-colonizing biomass detachment and determination, evaluating detachment efficiency and the effects of ultrasound exposure on bacterial cell viability. GAC samples were collected from two filters fed with groundwater. Conventional heterotrophic plate count (HPC) and fluorescence microscopy with a double staining method were used to evaluate cell viability, comparing two LES procedures, without and with periodical bulk substitution. A 20 min LES treatment, with bulk substitution after cycles of 5 min as maximum treatment time, allowed to recover 87%/100% of attached biomass, protecting detached bacteria from ultrasound damaging effects. Observed viable cell inactivation rate was 6.5/7.9% cell/min, with membrane-compromised cell damage appearing to be even higher (11.5%/13.1% cell/min). Assessing bacterial detachment and damaging ultrasound effects, fluorescence microscopy turned out to be more sensitive compared to conventional HPC. The optimized method revealed a GAC-colonizing biomass of 9.9 x 10(7) cell/gGAC for plant 1 and 8.8 x 10(7) cell/gGAC for plant 2, 2 log lower than reported in literature. The difference between the two GAC-colonizing biomasses is higher in terms of viable cells (46.3% of total cells in plant 1 GAC-colonizing biomass compared to the 33.3% in plant 2). Studying influent water contamination through multivariate statistical analyses, apossible combined toxic and genotoxic effect of chromium VI and trichloroethylene was suggested as a reason for the lower viable cell fraction observed in plant 2 GAC-colonizing population.

The Effects of Transient Retinal Detachment on Cavity Size and Glial and Neural Remodeling in a Mouse Model of X-Linked Retinoschisis

PubMed Central

Luna, Gabriel; Kjellstrom, Sten; Verardo, Mark R.; Lewis, Geoffrey P.; Byun, Jiyun; Sieving, Paul A.; Fisher, Steven K.

2009-01-01

Purpose To determine the cellular consequences of retinal detachment in retinoschisin knockout (Rs1-KO) mice, a model for retinoschisin in humans. Methods Experimental retinal detachments (RDs) were induced in the right eyes of both Rs1-KO and wild-type (wt) control mice. Immunocytochemistry was performed on retinal tissue at 1, 7, or 28 days after RD with antibodies to anti-GFAP, -neurofilament, and -rod opsin to examine cellular changes after detachment. Images of the immunostained tissue were captured by laser scanning confocal microscopy. Quantitative analysis was performed to measure the number of Hoechst-stained photoreceptor nuclei and their density, number, and size of inner retinal cavities, as well as the number of subretinal glial scars. Results Since detachments were created with balanced salt solution, by examination, all retinas had spontaneously reattached by 1 day. Cellular responses common to many photoreceptor degenerations occurred in the nondetached retinas of Rs1-KO mice, and, of importance, RD did not appear to significantly accentuate these responses. The number of schisis cavities was not changed after detachment, but their size was reduced. Conclusions These data indicate that large short-term RD in Rs1-KO mice, followed by a period of reattachment may cause a slight increase in photoreceptor cell death, but detachments do not accentuate the gliosis and neurite sprouting already present and may in fact reduce the size of existing retinal cavities. This finding suggests that performing subretinal injections to deliver therapeutic agents may be a viable option in the treatment of patients with retinoschisis without causing significant cellular damage to the retina. PMID:19387072

Evolution of Cell Size Homeostasis and Growth Rate Diversity during Initial Surface Colonization of Shewanella oneidensis.

PubMed

Lee, Calvin K; Kim, Alexander J; Santos, Giancarlo S; Lai, Peter Y; Lee, Stella Y; Qiao, David F; Anda, Jaime De; Young, Thomas D; Chen, Yujie; Rowe, Annette R; Nealson, Kenneth H; Weiss, Paul S; Wong, Gerard C L

2016-09-06

Cell size control and homeostasis are fundamental features of bacterial metabolism. Recent work suggests that cells add a constant size between birth and division ("adder" model). However, it is not known how cell size homeostasis is influenced by the existence of heterogeneous microenvironments, such as those during biofilm formation. Shewanella oneidensis MR-1 can use diverse energy sources on a range of surfaces via extracellular electron transport (EET), which can impact growth, metabolism, and size diversity. Here, we track bacterial surface communities at single-cell resolution to show that not only do bacterial motility appendages influence the transition from two- to three-dimensional biofilm growth and control postdivisional cell fates, they strongly impact cell size homeostasis. For every generation, we find that the average growth rate for cells that stay on the surface and continue to divide (nondetaching population) and that for cells that detach before their next division (detaching population) are roughly constant. However, the growth rate distribution is narrow for the nondetaching population, but broad for the detaching population in each generation. Interestingly, the appendage deletion mutants (ΔpilA, ΔmshA-D, Δflg) have significantly broader growth rate distributions than that of the wild type for both detaching and nondetaching populations, which suggests that Shewanella appendages are important for sensing and integrating environmental inputs that contribute to size homeostasis. Moreover, our results suggest multiplexing of appendages for sensing and motility functions contributes to cell size dysregulation. These results can potentially provide a framework for generating metabolic diversity in S. oneidensis populations to optimize EET in heterogeneous environments.

SU-F-T-678: Clotrimazole Sensitizes MCF-7 Breast Cancer Cell Line to Radiation

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

Garcia, L; Tambasco, M

2016-06-15

Purpose: To study the effects of Clotrimazole (CLT) on radiosensitivity of MCF-7 Cells in correlation to detachment of Hexokinase II from the Voltage Dependent Anion Channel on the outer membrane of the mitochondria. Apoptotic fractions were also analyzed in relation to the detachment of Hexokinase. Methods: This study focused on the mammary adenocarcinoma cell line, MCF-7. Colony forming assays were used to analyze radiosensitization by CLT. Flow cytometry methods were used to analyze apoptotic vs necrotic fractions after treatment with CLT. Spectrophotometery was used to analyze the mitochondrial bound and soluble fraction of Hexokinase by means of relative enzymatic activity.more » Results: Our preliminary data have shown that CLT sensitizes MCF-7 cells to radiation in a dose and incubation time dependent manner up. We have also demonstrated that there are two radiosensitizing periods in MCF-7 cells with the first corresponding to the cycle arrest after 24 hours observed in other cell lines. The second radiosensitizing period occurs with incubation in CLT after irradiation which reaches maximum effect around 24 hours of incubation time. Preliminary data from our Hexokinase detachment assay show a factor of two increase in the ratio of unbound to bound Hexokinase when comparing incubation for 24 hours in media containing 0 and 20 µM CLT. Conclusion: This study and others indicate CLT as a possible radiosensitizing agent in cancer therapies. While CLT itself shows toxicity to the liver in high doses, this study further demonstrates that disruption of the Warburg Effect and unbinding of mitochondrial bound Hexokinase as a possible pathway for cancer treatment.« less

A Review of Cell Adhesion Studies for Biomedical and Biological Applications.

PubMed

Khalili, Amelia Ahmad; Ahmad, Mohd Ridzuan

2015-08-05

Cell adhesion is essential in cell communication and regulation, and is of fundamental importance in the development and maintenance of tissues. The mechanical interactions between a cell and its extracellular matrix (ECM) can influence and control cell behavior and function. The essential function of cell adhesion has created tremendous interests in developing methods for measuring and studying cell adhesion properties. The study of cell adhesion could be categorized into cell adhesion attachment and detachment events. The study of cell adhesion has been widely explored via both events for many important purposes in cellular biology, biomedical, and engineering fields. Cell adhesion attachment and detachment events could be further grouped into the cell population and single cell approach. Various techniques to measure cell adhesion have been applied to many fields of study in order to gain understanding of cell signaling pathways, biomaterial studies for implantable sensors, artificial bone and tooth replacement, the development of tissue-on-a-chip and organ-on-a-chip in tissue engineering, the effects of biochemical treatments and environmental stimuli to the cell adhesion, the potential of drug treatments, cancer metastasis study, and the determination of the adhesion properties of normal and cancerous cells. This review discussed the overview of the available methods to study cell adhesion through attachment and detachment events.

A Review of Cell Adhesion Studies for Biomedical and Biological Applications

PubMed Central

Ahmad Khalili, Amelia; Ahmad, Mohd Ridzuan

2015-01-01

Cell adhesion is essential in cell communication and regulation, and is of fundamental importance in the development and maintenance of tissues. The mechanical interactions between a cell and its extracellular matrix (ECM) can influence and control cell behavior and function. The essential function of cell adhesion has created tremendous interests in developing methods for measuring and studying cell adhesion properties. The study of cell adhesion could be categorized into cell adhesion attachment and detachment events. The study of cell adhesion has been widely explored via both events for many important purposes in cellular biology, biomedical, and engineering fields. Cell adhesion attachment and detachment events could be further grouped into the cell population and single cell approach. Various techniques to measure cell adhesion have been applied to many fields of study in order to gain understanding of cell signaling pathways, biomaterial studies for implantable sensors, artificial bone and tooth replacement, the development of tissue-on-a-chip and organ-on-a-chip in tissue engineering, the effects of biochemical treatments and environmental stimuli to the cell adhesion, the potential of drug treatments, cancer metastasis study, and the determination of the adhesion properties of normal and cancerous cells. This review discussed the overview of the available methods to study cell adhesion through attachment and detachment events. PMID:26251901

Anoikis evasion in inflammatory breast cancer cells is mediated by Bim-EL sequestration

PubMed Central

Buchheit, C L; Angarola, B L; Steiner, A; Weigel, K J; Schafer, Z T

2015-01-01

Inflammatory breast cancer (IBC) is a rare and highly invasive type of breast cancer, and patients diagnosed with IBC often face a very poor prognosis. IBC is characterized by the lack of primary tumor formation and the rapid accumulation of cancerous epithelial cells in the dermal lymphatic vessels. Given that normal epithelial cells require attachment to the extracellular matrix (ECM) for survival, a comprehensive examination of the molecular mechanisms underlying IBC cell survival in the lymphatic vessels is of paramount importance to our understanding of IBC pathogenesis. Here we demonstrate that, in contrast to normal mammary epithelial cells, IBC cells evade ECM-detachment-induced apoptosis (anoikis). ErbB2 and EGFR knockdown in KPL-4 and SUM149 cells, respectively, causes decreased colony growth in soft agar and increased caspase activation following ECM detachment. ERK/MAPK signaling was found to operate downstream of ErbB2 and EGFR to protect cells from anoikis by facilitating the formation of a protein complex containing Bim-EL, LC8, and Beclin-1. This complex forms as a result of Bim-EL phosphorylation on serine 59, and thus Bim-EL cannot localize to the mitochondria and cause anoikis. These results reveal a novel mechanism that could be targeted with innovative therapeutics to induce anoikis in IBC cells. PMID:25526094

Anoikis evasion in inflammatory breast cancer cells is mediated by Bim-EL sequestration.

PubMed

Buchheit, C L; Angarola, B L; Steiner, A; Weigel, K J; Schafer, Z T

2015-08-01

Inflammatory breast cancer (IBC) is a rare and highly invasive type of breast cancer, and patients diagnosed with IBC often face a very poor prognosis. IBC is characterized by the lack of primary tumor formation and the rapid accumulation of cancerous epithelial cells in the dermal lymphatic vessels. Given that normal epithelial cells require attachment to the extracellular matrix (ECM) for survival, a comprehensive examination of the molecular mechanisms underlying IBC cell survival in the lymphatic vessels is of paramount importance to our understanding of IBC pathogenesis. Here we demonstrate that, in contrast to normal mammary epithelial cells, IBC cells evade ECM-detachment-induced apoptosis (anoikis). ErbB2 and EGFR knockdown in KPL-4 and SUM149 cells, respectively, causes decreased colony growth in soft agar and increased caspase activation following ECM detachment. ERK/MAPK signaling was found to operate downstream of ErbB2 and EGFR to protect cells from anoikis by facilitating the formation of a protein complex containing Bim-EL, LC8, and Beclin-1. This complex forms as a result of Bim-EL phosphorylation on serine 59, and thus Bim-EL cannot localize to the mitochondria and cause anoikis. These results reveal a novel mechanism that could be targeted with innovative therapeutics to induce anoikis in IBC cells.

Developing a Continuous Bioprocessing Approach to Stromal Cell Manufacture.

PubMed

Miotto, Martina; Gouveia, Ricardo; Abidin, Fadhilah Zainal; Figueiredo, Francisco; Connon, Che J

2017-11-29

To this day, the concept of continuous bioprocessing has been applied mostly to the manufacture of molecular biologics such as proteins, growth factors, and secondary metabolites with biopharmaceutical uses. The present work now sets to explore the potential application of continuous bioprocess methods to source large numbers of human adherent cells with potential therapeutic value. To this purpose, we developed a smart multifunctional surface coating capable of controlling the attachment, proliferation, and subsequent self-detachment of human corneal stromal cells. This system allowed the maintenance of cell cultures under steady-state growth conditions, where self-detaching cells were continuously replenished by the proliferation of those remaining attached. This facilitated a closed, continuous bioprocessing platform with recovery of approximately 1% of the total adherent cells per hour, a yield rate that was maintained for 1 month. Moreover, both attached and self-detached cells were shown to retain their original phenotype. Together, these results represent the proof-of-concept for a new high-throughput, high-standard, and low-cost biomanufacturing strategy with multiple potentials and important downstream applications.

Loss of anchorage primarily induces non-apoptotic cell death in a human mammary epithelial cell line under atypical focal adhesion kinase signaling.

PubMed

Ishikawa, F; Ushida, K; Mori, K; Shibanuma, M

2015-01-22

Anchorage dependence of cellular growth and survival prevents inappropriate cell growth or survival in ectopic environments, and serves as a potential barrier to metastasis of cancer cells. Therefore, obtaining a better understanding of anchorage-dependent responses in normal cells is the first step to understand and impede anchorage independence of growth and survival in cancer cells and finally to eradicate cancer cells during metastasis. Anoikis, a type of apoptosis specifically induced by lack of appropriate cell-extracellular matrix adhesion, has been established as the dominant response of normal epithelial cells to anchorage loss. For example, under detached conditions, the untransformed mammary epithelial cell (MEC) line MCF-10 A, which exhibits myoepithelial characteristics, underwent anoikis dependent on classical ERK signaling. On the other hand, recent studies have revealed a variety of phenotypes resulting in cell death modalities distinct from anoikis, such as autophagy, necrosis, and cornification, in detached epithelial cells. In the present study, we characterized detachment-induced cell death (DICD) in primary human MECs immortalized with hTERT ((Tert)HMECs), which are bipotent progenitor-like cells with a differentiating phenotype to luminal cells. In contrast to MCF-10 A cells, apoptosis was not observed in detached (Tert)HMECs; instead, non-apoptotic cell death marked by features of entosis, cornification, and necrosis was observed along with downregulation of focal adhesion kinase (FAK) signaling. Cell death was overcome by anchorage-independent activities of FAK but not PI3K/AKT, SRC, and MEK/ERK, suggesting critical roles of atypical FAK signaling pathways in the regulation of non-apoptotic cell death. Further analysis revealed an important role of TRAIL (tumor necrosis factor (TNF)-related apoptosis-inducing ligand) as a mediator of FAK signaling in regulation of entosis and necrosis and a role of p38 MAPK in the induction of necrosis. Overall, the present study highlighted outstanding cell subtype or differentiation stage specificity in cell death phenotypes induced upon anchorage loss in human MECs.

Morus alba Leaf Lectin (MLL) Sensitizes MCF-7 Cells to Anoikis by Inhibiting Fibronectin Mediated Integrin-FAK Signaling through Ras and Activation of P38 MAPK

PubMed Central

Saranya, Jayaram; Shilpa, Ganesan; Raghu, Kozhiparambil G.; Priya, Sulochana

2017-01-01

Lectins are a unique class of carbohydrate binding proteins/glycoproteins, and many of them possess anticancer properties. They can induce cell cycle arrest and apoptosis, inhibit protein synthesis, telomerase activity and angiogenesis in cancer cells. In the present study, we have demonstrated the effect of Morus alba leaf lectin (MLL) on anoikis induction in MCF-7 cells. Anoikis induction in cancer cells has a significant role in preventing early stage metastasis. MLL treatment in monolayers of MCF-7 cells caused significant detachment of cells in a time and concentration dependent manner. The detached cells failed to re-adhere and grew even to culture plates coated with different matrix proteins. DNA fragmentation, membrane integrity studies, annexin V staining, caspase 9 activation and upregulation of Bax/Bad confirmed that the detached cells underwent apoptosis. Upregulation of matrix metalloproteinase 9 (MMP-9) caused a decrease in fibronectin (FN) production which facilitated the cells to detach by blocking the FN mediated downstream signaling. On treatment with MLL, we have observed downregulation of integrin expression, decreased phosphorylation of focal adhesion kinase (FAK), loss in FAK-integrin interaction and active Ras. MLL treatment downregulated the levels of phosphorylated Akt and PI3K. Also, we have studied the effect of MLL on two stress activated protein kinases p38 MAPK and JNK. p38 MAPK activation was found to be elevated, but there was no change in the level of JNK. Thus our study substantiated the possible antimetastatic effect of MLL by inducing anoikis in MCF-7 cells by activation of caspase 9 and proapoptotic Bax/Bad by blockage of FN mediated integrin/FAK signaling and partly by activation of p38 MAPK. PMID:28223935

Microscopic Behavior Of Colloidal Particles Under The Effect Of Acoustic Stimulations In The Ultrasonic To Megasonic Range

NASA Astrophysics Data System (ADS)

Abdel-Fattah, Amr I.; Roberts, Peter M.

2006-05-01

It is well known that colloid attachment and detachment at solid surfaces are influenced strongly by physico-chemical conditions controlling electric double layer (EDL) and solvation-layer effects. We present experimental observations demonstrating that, in addition, acoustic waves can produce strong effects on colloid/surface interactions that can alter the behavior of colloid and fluid transport in porous media. Microscopic colloid visualization experiments were performed with polystyrene micro-spheres suspended in water in a parallel-plate glass flow cell. When acoustic energy was applied to the cell at frequencies from 500 kHz to 5 MHz, changes in colloid attachment to and detachment from the glass cell surfaces were observed. Quantitative measurements of acoustically-induced detachment of 300-nm microspheres in 0.1M NaCl solution demonstrated that roughly 30% of the colloids that were attached to the glass cell wall during flow alone could be detached rapidly by applying acoustics at frequencies in the range of 0.7 to 1.2 MHz. The remaining attached colloids could not be detached by acoustics. This implies the existence of both "strong" and "weak" attachment sites at the cell surface. Subsequent re-attachment of colloids with acoustics turned off occurred only at new, previously unoccupied sites. Thus, acoustics appears to accelerate simultaneously both the deactivation of existing weak sites where colloids are already attached, and the activation of new weak sites where future attachments can occur. Our observations indicate that acoustics (and, in general, dynamic stress) can influence colloid-colloid and colloid-surface interactions in ways that could cause significant changes in porous-media permeability and mass transport. This would occur due to either buildup or release of colloids present in the porous matrix.

Coupled factors influencing detachment of nano- and micro-sized particles from primary minima.

PubMed

Shen, Chongyang; Lazouskaya, Volha; Jin, Yan; Li, Baoguo; Ma, Zhiqiang; Zheng, Wenjuan; Huang, Yuanfang

2012-06-01

This study examined the detachments of nano- and micro-sized colloids from primary minima in the presence of cation exchange by laboratory column experiments. Colloids were initially deposited in columns packed with glass beads at 0.2 M CaCl(2) in the primary minima of Derjaguin-Landau-Verwey-Overbeek (DLVO) interaction energies. Then, the columns were flushed with NaCl solutions with different ionic strengths (i.e., 0.001, 0.01, 0.1 and 0.2 M). Detachments were observed at all ionic strengths and were particularly significant for the nanoparticle. The detachments increased with increasing electrolyte concentration for the nanoparticle whereas increased from 0.001 M to 0.01 M and decreased with further increasing electrolyte concentration for the micro-sized colloid. The observations were attributed to coupled influence of cation exchange, short-range repulsion, surface roughness, surface charge heterogeneity, and deposition in the secondary minima. The detachments of colloids from primary minima challenge the common belief that colloid interaction in primary minimum is irreversible and resistant to disturbance in solution ionic strength and composition. Although the significance of surface roughness, surface charge heterogeneity, and secondary minima on colloid deposition has been widely recognized, our study implies that they also play important roles in colloid detachment. Whereas colloid detachment is frequently associated with decrease of ionic strength, our results show that increase of ionic strength can also cause detachment due to influence of cation exchange. Copyright © 2012 Elsevier B.V. All rights reserved.

Impact of biofilm formation and detachment on the transmission of bacterial antibiotic resistance in drinking water distribution systems.

PubMed

Zhang, Junpeng; Li, Weiying; Chen, Jiping; Qi, Wanqi; Wang, Feng; Zhou, Yanyan

2018-07-01

There is growing awareness of the antibiotic-resistance crisis and its implications for public health among clinicians, researchers, politicians, and the public. We studied bacterial antibiotic resistance transition and the role of biofilms in a drinking water distribution system (DWDS). We tracked several different antibiotic resistant bacteria (ARB) with resistance to tetracycline, sulfamethoxazole, clindamycin, and norfloxacin for one year in a DWDS. The results indicated that the amount of ARB increased in tap water, presumably due to biofilm detachment. The effect of biofilm detachment on the transmission of antibiotic resistance from biofilms to tap water was explored by using a bacterial annular reactor. The percentage of ARB of inlet water, outlet water, and biofilms ranged from 0.26% to 9.85%, 1.08%-16.29%, and 0.52%-29.97%, respectively in a chlorinated system, and from 0.23% to 9.89%, 0.84%-16.84%, and 0.35%-17.77%, respectively, in a chloraminated system. The relative abundances of antibiotic resistance Acinetobacter, Sphingomonas, and Bradyrhizobium were higher in outlet water than in inlet water, as determined by high throughout sequencing. The amount of ARB percentage varied with the concentration of viable but non-culturable (VBNC) cells (r = 0.21, n = 160, P < 0.05) in biofilm, suggesting a higher antibiotic resistance mutation rate in VBNC cells. Our results suggest that biofilm detachment was promoted by disinfectant and affected the overall bacterial antibiotic resistance of microbes in tap water. Copyright © 2018 Elsevier Ltd. All rights reserved.

Effects of low-Z and high-Z impurities on divertor detachment and plasma confinement

DOE PAGES

Wang, H. Q.; Guo, Houyang Y.; Petrie, Thomas W.; ...

2017-03-18

The impurity-seeded detached divertor is essential for heat exhaust in ITER and other reactor-relevant devices. Dedicated experiments with injection of N 2, Ne and Ar have been performed in DIII-D to assess the impact of the different impurities on divertor detachment and confinement. Seeding with N 2, Ne and Ar all promote divertor detachment, greatly reducing heat flux near the strike point. The upstream plasma density at the onset of detachment decreases with increasing impurity-puffing flow rates. For all injected impurity species, the confinement and pedestal pressure are correlated with the impurity content and the ratio of separatrix loss powermore » to the L-H transition threshold power. As the divertor plasma approaches detachment, the high-Z impurity seeding tends to degrade the core confinement owing to the increased core radiation. In particular, Ar injection leads to an increase in core radiation, up to 50% of the injected power, and a reduction in pedestal temperature over 60%, thus significantly degrading the confinement, i.e., with H 98 reducing from 1.1 to below 0.7. As for Ne seeding, H 98 near 0.8 can be maintained during the detachment phase with the pedestal temperature being reduced by about 50%. In contrast, in the N 2 seeded plasmas, radiation is predominately confined in the boundary plasma, with up to 50% of heating power being radiated in the divertor region and less than 25% in the core at the onset of detachment. In the case of strong N 2 gas puffing, the confinement recovers during the detachment, from ~20% reduction at the onset of the detachment to greater than that before the seeding. The core and pedestal temperatures feature a reduction of 30% from the initial attached phase and remain nearly constant during the detachment phase. The improvement in confinement appears to arise from the increase in pedestal and core density despite the temperature reduction.« less

Human dental pulp stem cell adhesion and detachment in polycaprolactone electrospun scaffolds under direct perfusion

PubMed Central

Paim, A.; Braghirolli, D.I.; Cardozo, N.S.M.; Pranke, P.; Tessaro, I.C.

2018-01-01

Cell adhesion in three-dimensional scaffolds plays a key role in tissue development. However, stem cell behavior in electrospun scaffolds under perfusion is not fully understood. Thus, an investigation was made on the effect of flow rate and shear stress, adhesion time, and seeding density under direct perfusion in polycaprolactone electrospun scaffolds on human dental pulp stem cell detachment. Polycaprolactone scaffolds were electrospun using a solvent mixture of chloroform and methanol. The viable cell number was determined at each tested condition. Cell morphology was analyzed by confocal microscopy after various incubation times for static cell adhesion with a high seeding density. Scanning electron microscopy images were obtained before and after perfusion for the highest flow rate tested. The wall pore shear stress was calculated for all tested flow rates (0.005–3 mL/min). An inversely proportional relationship between adhesion time with cell detachment under perfusion was observed. Lower flow rates and lower seeding densities reduced the drag of cells by shear stress. However, there was an operational limit for the lowest flow rate that can be used without compromising cell viability, indicating that a flow rate of 0.05 mL/min might be more suitable for the tested cell culture in electrospun scaffolds under direct perfusion. PMID:29590258

A Role for MEK-Interacting Protein 1 In Hormone Responsiveness of ER Positive Breast Cancer Cells

DTIC Science & Technology

2011-10-01

48 hours, ER- positiv e cell lines tran sfected with MP1siRNA (but not control siR NA) rounded up and detached fr om the plate, and trypan blue...phenotype to MCF-7. To quantitate the effect of MP1 knockdown, attached and detached cells were collected at 48 h following siRNA transfection, stained...Immunoblot from a representative experiment. Lower panel: Quantitation of MP1/Actin ratios in three independent experiments (mean ± SD, *pɘ.05). Figure

Mechanism by Which Cold Shock Evokes Exocytosis of Symbiotic Algae in Marine Cnidarians

DTIC Science & Technology

1993-05-30

shock, and UV irradiation, and the mode of release of zooxanthellae was investigated. It was determined that all three types of acute stress evoked...detachment and expulsion of intact host endoderm cells containing zooxanthellae . To determine how cold shock evokes cell detachment we have formulated...Coral Reefs 11:143-154. Bil’, K., P. Kolmakov. and L. Muscatine (1992) Photosynthetic products of zooxanthellae of the reef building corals

Near-IR laser-triggered target cell collection using a carbon nanotube-based cell-cultured substrate.

PubMed

Sada, Takao; Fujigaya, Tsuyohiko; Niidome, Yasuro; Nakazawa, Kohji; Nakashima, Naotoshi

2011-06-28

Unique near-IR optical properties of single-walled carbon nanotube (SWNTs) are of interest in many biological applications. Here we describe the selective cell detachment and collection from an SWNT-coated cell-culture dish triggered by near-IR pulse laser irradiation. First, HeLa cells were cultured on an SWNT-coated dish prepared by a spraying of an aqueous SWNT dispersion on a glass dish. The SWNT-coated dish was found to show a good cell adhesion behavior as well as a cellular proliferation rate similar to a conventional glass dish. We discovered, by near-IR pulse laser irradiation (at the laser power over 25 mW) to the cell under optical microscopic observation, a quick single-cell detachment from the SWNT-coated surface. Shockwave generation from the irradiated SWNTs is expected to play an important role for the cell detachment. Moreover, we have succeeded in catapulting the target single cell from the cultured medium when the depth of the medium was below 150 μm and the laser power was stronger than 40 mW. The captured cell maintained its original shape. The retention of the genetic information of the cell was confirmed by the polymerase chain reaction (PCR) technique. A target single-cell collection from a culture medium under optical microscopic observation is significant in wide fields of single-cell studies in biological areas.

Exposure to Bordetella pertussis adenylate cyclase toxin affects integrin-mediated adhesion and mechanics in alveolar epithelial cells.

PubMed

Angely, Christelle; Nguyen, Ngoc-Minh; Andre Dias, Sofia; Planus, Emmanuelle; Pelle, Gabriel; Louis, Bruno; Filoche, Marcel; Chenal, Alexandre; Ladant, Daniel; Isabey, Daniel

2017-08-01

The adenylate cyclase (CyaA) toxin is a major virulent factor of Bordetella pertussis, the causative agent of whooping cough. CyaA toxin is able to invade eukaryotic cells where it produces high levels of cyclic adenosine monophosphate (cAMP) affecting cellular physiology. Whether CyaA toxin can modulate cell matrix adhesion and mechanics of infected cells remains largely unknown. In this study, we use a recently proposed multiple bond force spectroscopy (MFS) with an atomic force microscope to assess the early phase of cell adhesion (maximal detachment and local rupture forces) and cell rigidity (Young's modulus) in alveolar epithelial cells (A549) for toxin exposure <1 h. At 30 min of exposure, CyaA toxin has a minimal effect on cell viability (>95%) at CyaA concentration of 0.5 nM, but a significant effect (≈81%) at 10 nM. MFS performed on A549 for three different concentrations (0.5, 5 and 10 nM) demonstrates that CyaA toxin significantly affects both cell adhesion (detachment forces are decreased) and cell mechanics (Young's modulus is increased). CyaA toxin (at 0.5 nM) assessed at three indentation/retraction speeds (2, 5 and 10 μm/s) significantly affects global detachment forces, local rupture events and Young modulus compared with control conditions, while an enzymatically inactive variant CyaAE5 has no effect. These results reveal the loading rate dependence of the multiple bonds newly formed between the cell and integrin-specific coated probe as well as the individual bond kinetics which are only slightly affected by the patho-physiological dose of CyaA toxin. Finally, theory of multiple bond force rupture enables us to deduce the bond number N which is reduced by a factor of 2 upon CyaA exposure (N ≈ 6 versus N ≈ 12 in control conditions). MFS measurements demonstrate that adhesion and mechanical properties of A549 are deeply affected by exposure to the CyaA toxin but not to an enzymatically inactive variant. This indicates that the alteration of cell mechanics triggered by CyaA is a consequence of the increase in intracellular cAMP in these target cells. These results suggest that mechanical and adhesion properties of the cells appear as pertinent markers of cytotoxicity of CyaA toxin. © 2017 Société Française des Microscopies and Société de Biologie Cellulaire de France. Published by John Wiley & Sons Ltd.

Thermo-responsive cell culture carrier: Effects on macrophage functionality and detachment efficiency.

PubMed

Rennert, Knut; Nitschke, Mirko; Wallert, Maria; Keune, Natalie; Raasch, Martin; Lorkowski, Stefan; Mosig, Alexander S

2017-01-01

Harvesting cultivated macrophages for tissue engineering purposes by enzymatic digestion of cell adhesion molecules can potentially result in unintended activation, altered function, or behavior of these cells. Thermo-responsive polymer is a promising tool that allows for gentle macrophage detachment without artificial activation prior to subculture within engineered tissue constructs. We therefore characterized different species of thermo-responsive polymers for their suitability as cell substrate and to mediate gentle macrophage detachment by temperature shift. Primary human monocyte- and THP-1-derived macrophages were cultured on thermo-responsive polymers and characterized for phagocytosis and cytokine secretion in response to lipopolysaccharide stimulation. We found that both cell types differentially respond in dependence of culture and stimulation on thermo-responsive polymers. In contrast to THP-1 macrophages, primary monocyte-derived macrophages showed no signs of impaired viability, artificial activation, or altered functionality due to culture on thermo-responsive polymers compared to conventional cell culture. Our study demonstrates that along with commercially available UpCell carriers, two other thermo-responsive polymers based on poly(vinyl methyl ether) blends are attractive candidates for differentiation and gentle detachment of primary monocyte-derived macrophages. In summary, we observed similar functionality and viability of primary monocyte-derived macrophages cultured on thermo-responsive polymers compared to standard cell culture surfaces. While this first generation of custom-made thermo-responsive polymers does not yet outperform standard culture approaches, our results are very promising and provide the basis for exploiting the unique advantages offered by custom-made thermo-responsive polymers to further improve macrophage culture and recovery in the future, including the covalent binding of signaling molecules and the reduction of centrifugation and washing steps. Optimizing these and other benefits of thermo-responsive polymers could greatly improve the culture of macrophages for tissue engineering applications.

Cell-matrix adhesion characterization using multiple shear stress zones in single stepwise microchannel

NASA Astrophysics Data System (ADS)

Kim, Min-Ji; Doh, Il; Bae, Gab-Yong; Cha, Hyuk-Jin; Cho, Young-Ho

2014-08-01

This paper presents a cell chip capable to characterize cell-matrix adhesion by monitoring cell detachment rate. The proposed cell chip can supply multiple levels of shear stress in single stepwise microchannel. As epithelial-mesenchymal transition (EMT), one of hallmarks of cancer metastasis is closely associated to the interaction with extracelluar matrix (ECM), we took advantage of two lung cancer cell models with different adhesion properties to ECM depending their epithelial or mesenchymal properties, including the pair of lung cancer cells with (A549sh) or without E-cadherin expression (A549sh-Ecad), which would be optimal model to examine the alteration of adhesion properties after EMT induction. The cell-matrix adhesion resisting to shear stress appeared to be remarkably differed between lung cancer cells. The detachment rate of epithelial-like H358 and mesenchymal-like H460 cells was 53%-80% and 25%-66% in the shear stress range of 34-60 dyn/cm2, respectively. A549sh-Ecad cells exhibits lower detachment rate (5%-9%) compared to A549sh cells (14%-40%). By direct comparison of adhesion between A549sh and A549sh-Ecad, we demonstrated that A549shE-cad to mimic EMT were more favorable to the ECM attachment under the various levels of shear stress. The present method can be applied to quantitative analysis of tumor cell-ECM adhesion.

Transfer of fibroblast sheets cultured on thermoresponsive dishes with membranes.

PubMed

Kawecki, Marek; Kraut, Małgorzata; Klama-Baryła, Agnieszka; Łabuś, Wojciech; Kitala, Diana; Nowak, Mariusz; Glik, Justyna; Sieroń, Aleksander L; Utrata-Wesołek, Alicja; Trzebicka, Barbara; Dworak, Andrzej; Szweda, Dawid

2016-06-01

In cell or tissue engineering, it is essential to develop a support for cell-to-cell adhesion, which leads to the generation of cell sheets connected by extracellular matrix. Such supports must be hydrophobic and should result in a detachable cell sheet. A thermoresponsive support that enables the cultured cell sheet to detach using only a change in temperature could be an interesting alternative in regenerative medicine. The aim of this study was to evaluate plates covered with thermoresponsive polymers as supports for the formation of fibroblast sheets and to develop a damage-free procedure for cell sheet transfer with the use of membranes as transfer tools. Human skin fibroblasts were seeded on supports coated with a thermoresponsive polymer: commercial UpCell™ dishes (NUNC™) coated with thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) and dishes coated with thermoresponsive poly(tri(ethylene glycol) monoethyl ether methacrylate) (P(TEGMA-EE)). Confluent fibroblast sheets were effectively cultured and harvested from both commercial PNIPAM-coated dishes and laboratory P(TEGMA-EE)-coated dishes. To transfer a detached cell sheet, two membranes, Immobilon-P(®) and SUPRATHEL(®), were examined. The use of SUPRATHEL for relocating the cell sheets opens a new possibility for the clinical treatment of wounds. This study established the background for implementing thermoresponsive supports for transplanting in vitro cultured fibroblasts.

Quantitative Fundus Autofluorescence in Best Vitelliform Macular Dystrophy: RPE Lipofuscin is not Increased in Non-Lesion Areas of Retina

PubMed Central

Duncker, Tobias; Woods, Russell; Delori, François C.

2018-01-01

Since the lipofuscin of retinal pigment epithelial (RPE) cells has been implicated in the pathogenesis of Best vitelliform macular dystrophy, we quantified fundus autofluorescence (quantitative fundus autofluorescence, qAF) as an indirect measure of RPE lipofuscin levels. Mean non-lesion qAF was found to be within normal limits for age. By spectral domain optical coherence tomography (SD-OCT) vitelliform lesions presented as fluid-filled subretinal detachments containing reflective material. We discuss photoreceptor outer segment debris as the source of the intense fluorescence of these lesions and loss of anion channel functioning as an explanation for the bullous photoreceptor-RPE detachment. Unexplained is the propensity of the disease for central retina. PMID:26427423

Quantitative Fundus Autofluorescence in Best Vitelliform Macular Dystrophy: RPE Lipofuscin is not Increased in Non-Lesion Areas of Retina.

PubMed

Sparrow, Janet R; Duncker, Tobias; Woods, Russell; Delori, François C

2016-01-01

Since the lipofuscin of retinal pigment epithelial (RPE) cells has been implicated in the pathogenesis of Best vitelliform macular dystrophy, we quantified fundus autofluorescence (quantitative fundus autofluorescence, qAF) as an indirect measure of RPE lipofuscin levels. Mean non-lesion qAF was found to be within normal limits for age. By spectral domain optical coherence tomography (SD-OCT) vitelliform lesions presented as fluid-filled subretinal detachments containing reflective material. We discuss photoreceptor outer segment debris as the source of the intense fluorescence of these lesions and loss of anion channel functioning as an explanation for the bullous photoreceptor-RPE detachment. Unexplained is the propensity of the disease for central retina.

Heat transport in bubbling turbulent convection

PubMed Central

Lakkaraju, Rajaram; Stevens, Richard J. A. M.; Oresta, Paolo; Verzicco, Roberto; Lohse, Detlef; Prosperetti, Andrea

2013-01-01

Boiling is an extremely effective way to promote heat transfer from a hot surface to a liquid due to numerous mechanisms, many of which are not understood in quantitative detail. An important component of the overall process is that the buoyancy of the bubble compounds with that of the liquid to give rise to a much-enhanced natural convection. In this article, we focus specifically on this enhancement and present a numerical study of the resulting two-phase Rayleigh–Bénard convection process in a cylindrical cell with a diameter equal to its height. We make no attempt to model other aspects of the boiling process such as bubble nucleation and detachment. The cell base and top are held at temperatures above and below the boiling point of the liquid, respectively. By keeping this difference constant, we study the effect of the liquid superheat in a Rayleigh number range that, in the absence of boiling, would be between 2 × 106 and 5 × 109. We find a considerable enhancement of the heat transfer and study its dependence on the number of bubbles, the degree of superheat of the hot cell bottom, and the Rayleigh number. The increased buoyancy provided by the bubbles leads to more energetic hot plumes detaching from the cell bottom, and the strength of the circulation in the cell is significantly increased. Our results are in general agreement with recent experiments on boiling Rayleigh–Bénard convection. PMID:23696657

Heat transport in bubbling turbulent convection.

PubMed

Lakkaraju, Rajaram; Stevens, Richard J A M; Oresta, Paolo; Verzicco, Roberto; Lohse, Detlef; Prosperetti, Andrea

2013-06-04

Boiling is an extremely effective way to promote heat transfer from a hot surface to a liquid due to numerous mechanisms, many of which are not understood in quantitative detail. An important component of the overall process is that the buoyancy of the bubble compounds with that of the liquid to give rise to a much-enhanced natural convection. In this article, we focus specifically on this enhancement and present a numerical study of the resulting two-phase Rayleigh-Bénard convection process in a cylindrical cell with a diameter equal to its height. We make no attempt to model other aspects of the boiling process such as bubble nucleation and detachment. The cell base and top are held at temperatures above and below the boiling point of the liquid, respectively. By keeping this difference constant, we study the effect of the liquid superheat in a Rayleigh number range that, in the absence of boiling, would be between 2 × 10(6) and 5 × 10(9). We find a considerable enhancement of the heat transfer and study its dependence on the number of bubbles, the degree of superheat of the hot cell bottom, and the Rayleigh number. The increased buoyancy provided by the bubbles leads to more energetic hot plumes detaching from the cell bottom, and the strength of the circulation in the cell is significantly increased. Our results are in general agreement with recent experiments on boiling Rayleigh-Bénard convection.

Biophysics of cancer progression and high-throughput mechanical characterization of biomaterials

NASA Astrophysics Data System (ADS)

Osborne, Lukas Dylan

Cancer metastasis involves a series of events known as the metastatic cascade. In this complex progression, cancer cells detach from the primary tumor, invade the surrounding stromal space, transmigrate the vascular system, and establish secondary tumors at distal sites. Specific mechanical phenotypes are likely adopted to enable cells to successfully navigate the mechanical environments encountered during metastasis. To examine the role of cell mechanics in cancer progression, I employed force-consistent biophysical and biochemical assays to characterize the mechanistic links between stiffness, stiffness response and cell invasion during the epithelial to mesenchymal transition (EMT). EMT is an essential physiological process, whose abnormal reactivation has been implicated in the detachment of cancer cells from epithelial tissue and their subsequent invasion into stromal tissue. I demonstrate that epithelial-state cells respond to force by evoking a stiffening response, and that after EMT, mesenchymal-state cells have reduced stiffness but also lose the ability to increase their stiffness in response to force. Using loss and gain of function studies, two proteins are established as functional connections between attenuated stiffness and stiffness response and the increased invasion capacity acquired after EMT. To enable larger scale assays to more fully explore the connection between biomechanics and cancer, I discuss the development of an automated array high throughput (AHT) microscope. The AHT system is shown to implement passive microbead rheology to accurately characterize the mechanical properties of biomaterials. Compared to manually performed mechanical characterizations, the AHT system executes experiments in two orders of magnitude less time. Finally, I use the AHT microscope to study the effect of gain of function oncogenic molecules on cell stiffness. I find evidence that our assay can identify alterations in cell stiffness due to constitutive activation of cancer pathways.

Pathogenesis of rhegmatogenous retinal detachment: predisposing anatomy and cell biology.

PubMed

Mitry, Danny; Fleck, Brian W; Wright, Alan F; Campbell, Harry; Charteris, David G

2010-01-01

The pathogenesis of rhegmatogenous retinal detachment is complex, and our knowledge of the exact mechanism of vitreoretinal attachment and detachment remains incomplete. We performed a Medline, Ovid, and EMBASE search using search words rhegmatogenous, retinal detachment, vitreous, and retinal adhesion. All appropriate articles were reviewed, and the evidence was compiled. Cortical vitreous contains fibrillar collagens type II, V/XI, and IX. The inner limiting membrane of the retina contains collagens type I, IV, VI, and XVIII as well as numerous other glycoproteins and potential adhesion molecules. The distribution and age-related changes in the structure of these molecules play an important role in the formation of a retinal break, which may compromise and disrupt the normal mechanisms of neurosensory retinal adhesion. Rhegmatogenous retinal detachment development is intimately related to changes in the fibrillar structure of the aging vitreous culminating in posterior vitreous detachment with regions of persistent and tangential vitreoretinal traction predisposing to retinal tear formation. A complex interplay of factors such as weakening of vitreoretinal adhesion, posterior migration of the vitreous base, and molecular changes at the vitreoretinal interface are important in predisposing to focal areas of vitreoretinal traction precipitating rhegmatogenous retinal detachment. Once formed, the passage of liquefied vitreous through a retinal break may overwhelm normal neurosensory-retinal pigment epithelium adhesion perpetuating and extending detachment and causing visual loss. To understand the molecular events underlying rhegmatogenous retinal detachment so that new therapies can be developed, it is important to appreciate the structural organization of the vitreous, the biology underlying vitreous liquefaction and posterior vitreous detachment, and the mechanisms of vitreoretinal attachment and detachment.

Autophagy promotes degradation of internalized collagen and regulates distribution of focal adhesions to suppress cell adhesion

PubMed Central

Kawano, Shinichi; Esaki, Motohiro; Torisu, Kumiko; Matsuno, Yuichi; Kitazono, Takanari

2017-01-01

ABSTRACT Adhesion of cells to the extracellular matrix (ECM) via focal adhesions (FAs) is crucial for cell survival, migration, and differentiation. Although the regulation of FAs, including by integrins and the ECM, is important to cell behavior, how FAs are regulated is not well known. Autophagy is induced by both cell adhesion and cell detachment. Here, we showed that autophagosomes are located close to internalized collagen and paxillin, which is a well-known marker of FAs. Autophagy-deficient cells showed increased levels of internalized collagen compared with control cells. Moreover, paxillin exhibited a more peripheral distribution and the area of paxillin was increased, and adhesion-induced focal adhesion kinase signaling was impaired and adhesion was enhanced, in autophagy-deficient cells. These results suggest that autophagy suppressed cell adhesion by regulating internalized ECM and FAs. PMID:28970230

Efficacy and mechanisms of non-antibacterial, chemical plaque control by dentifrices--an in vitro study.

PubMed

Busscher, Henk J; White, Don J; Atema-Smit, Jelly; van der Mei, Henny C

2007-04-01

The provision of antiplaque benefits to dentifrices assists patients in improving hygiene and reducing susceptibility to gingivitis and caries. Chemical plaque control involves different mechanisms and is mostly associated with antibacterial effects, but also includes effects on pellicle surface chemistry to improve cleansing or discourage renewed plaque formation. It is the aim of this paper to analyze in vitro detachment of co-aggregating oral actinomyces and streptococci from pellicle surfaces by dentifrice supernates and to study subsequent de novo streptococcal deposition. Detachment by dentifrices of a co-adhering bacterial pair was studied in the parallel plate flow chamber on a 16 h pellicle coated surface. After detachment by perfusing the chamber with a dentifrice, re-deposition was initiated by flowing with a fresh streptococcal suspension. The dentifrices included both a regular, SLS-fluoride based formulation as well a pyrophosphate, anticalculus and antimicrobial formulations. All dentifrice supernates containing SLS were effective in detaching co-adhering bacteria from pellicles surfaces, except in combination with SnF(2). When hexametaphosphate was added immediate detachment was relatively low, but continued even during re-deposition. The re-deposition of streptococci after detachment by other, NaF containing dentifrices involved relatively few large aggregates, presumably because fluoride was able to block bi-dentate calcium binding sites on the bacterial cell surfaces, mediating co-adhesion. When pyrophosphate was present in addition to NaF, re-deposition involved significantly more large aggregates, likely because pyrophosphate served as a bi-dentate bridge between calcium bound on the bacterial cell surfaces. Commercially available dentifrice formulations differ in their ability to stimulate bacterial detachment from pellicles and dependent on their composition yield the formation of large co-adhering aggregates of actinomyces and streptococci in de novo deposition.

Opposite effects of flurbiprofen and the nitroxybutyl ester of flurbiprofen on apoptosis in cultured guinea-pig gastric mucous cells

PubMed Central

Johal, Kamaljit; Hanson, Peter J

2000-01-01

The nitric oxide (NO)-donating nitroxybutyl ester of flurbiprofen (NO-flurbiprofen), shows reduced gastro-intestinal toxicity relative to flurbiprofen. NO may exert either pro- or anti-apoptotic effects, while non-steroidal anti-inflammatory drugs may induce apoptosis. The aim of the present work was therefore to compare the effects of flurbiprofen and NO-flurbiprofen on apoptosis in guinea-pig gastric mucous cells. Apoptotic activity was assessed by assay of caspase activity and from the fragmentation and condensation of nuclei. Incubation with flurbiprofen for 24 h produced a concentration-dependent induction of apoptosis in cells attached to the culture plate (caspase 3-like activity increased by 257% at 500 μM), while NO-flurbiprofen inhibited basal apoptosis (caspase 3-like activity decreased by 71% at 500 μM). Caspase activity and nuclear fragmentation were substantially increased in cells that had spontaneously detached from the culture plate. NO-flurbiprofen inhibited caspase activity (55% at 500 μM) but not nuclear fragmentation in these detached cells. NO flurbiprofen inhibited the activation of apoptosis by 25 μM C6-ceramide in cells attached to the culture plate. Inhibition of caspase activity by NO-flurbiprofen was detectable after 6 h of incubation with intact cells, but by contrast with the NO-donor S-nitrosyl-N-acetyl-penicillamine, was not demonstrable with cell homogenates. Activation of caspase 3-like activity by flurbiprofen was slow (>6 h incubation needed) and was inhibited by cycloheximide. The presence of a nitroxybutyl ester moiety on flurbiprofen prevents the pro-apoptotic activity of the parent compound and may contribute to the reduced gastro-intestinal toxicity of NO-flurbiprofen. PMID:10864887

Synchronous induction of detachment and reattachment of symbiotic Chlorella spp. from the cell cortex of the host Paramecium bursaria.

PubMed

Kodama, Yuuki; Fujishima, Masahiro

2013-09-01

Paramecium bursaria harbor several hundred symbiotic Chlorella spp. Each alga is enclosed in a perialgal vacuole membrane, which can attach to the host cell cortex. How the perialgal vacuole attaches beneath the host cell cortex remains unknown. High-speed centrifugation (> 1000×g) for 1min induces rapid detachment of the algae from the host cell cortex and concentrates the algae to the posterior half of the host cell. Simultaneously, most of the host acidosomes and lysosomes accumulate in the anterior half of the host cell. Both the detached algae and the dislocated acidic vesicles recover their original positions by host cyclosis within 10min after centrifugation. These recoveries were inhibited if the host cytoplasmic streaming was arrested by nocodazole. Endosymbiotic algae during the early reinfection process also show the capability of desorption after centrifugation. These results demonstrate that adhesion of the perialgal vacuole beneath the host cell cortex is repeatedly inducible, and that host cytoplasmic streaming facilitates recovery of the algal attachment. This study is the first report to illuminate the mechanism of the induction to desorb for symbiotic algae and acidic vesicles, and will contribute to the understanding of the mechanism of algal and organelle arrangements in Paramecium. Copyright © 2013 Elsevier GmbH. All rights reserved.

The nature of thrombosis induced by platinum and tungsten coils in saccular aneurysms.

PubMed

Byrne, J V; Hope, J K; Hubbard, N; Morris, J H

1997-01-01

To compare the efficacy and biocompatability of electrolytic and mechanically detachable embolization coils of two metal types. Experimental saccular aneurysms in pigs were used to assess embolization induced by platinum or tungsten coils. Longitudinal angiographic and histologic studies were performed on treated and untreated (control) aneurysms to compare thrombosis and cellular responses after embolization with electrolytically detachable platinum coils and with mechanically detached tungsten coils. Fewer tungsten than platinum coils were needed to induce thrombosis. The inflammatory response within the aneurysmal lumen was more florid in embolized aneurysms than in control aneurysms. No difference was found in the timing or extent of accumulation of eosinophils, lymphocytes, or polymorphs between the two coils used. Giant cell responses were more marked in treated aneurysms; tungsten coils more than platinum coils. The amount of collagen and fibrosis present increased over the study period and was similar in treated and control aneurysms. The coil type influenced the initial cellular response but had little effect on the rate or degree to which blood clot within the aneurysm was replaced by fibrous tissue.

Impact of Cross-field Drifts on Detachment in DIII-D

NASA Astrophysics Data System (ADS)

Jaervinen, A. E.; Allen, S. L.; McLean, A. G.; Rognlien, T. D.; Samuell, C. M.; Porter, G. D.; Groth, M.; Hill, D. N.; Leonard, A. W.

2017-10-01

Simulations of DIII-D plasmas have revealed the strong role of E ×B-drifts in the low field side (LFS) detachment structure. High confinement modes (H-mode) with the ∇B-drift towards the X-point (fwd BT) enter detachment at 20% higher upstream density, ne,sep, than plasmas with the ∇B-drift away from the X-point (rev BT). In contrast, low confinement modes (L-mode) enter detachment at 10% lower ne,sep in fwd BT. Despite this, both L- and H-modes detached plasmas show strong target flux, JSAT, reduction with increasing ne,sep in fwd BT, while only a modest reduction occurs in rev BT. In fwd BT H-mode, a step-wise transition from attached to strongly detached conditions is observed with increasing ne,sep. UEDGE simulations indicate that the strong poloidal E ×B-drift in the private flux region in H-mode drives the difference for the detachment onset relative to L-mode. In fwd BT, the dependence of this poloidal E ×B-drift on the divertor conditions can reinforce the plasma into either attached or strongly detached state. In rev BT, radial E ×B-drift depletes strike-line ne, limiting the degree of detachment. Work supported by the US DOE under DE-FC02-04ER54698, DE-AC52-07NA27344, and LLNL LDRD project 17-ERD-020.

P38 delta MAPK promotes breast cancer progression and lung metastasis by enhancing cell proliferation and cell detachment.

PubMed

Wada, M; Canals, D; Adada, M; Coant, N; Salama, M F; Helke, K L; Arthur, J S; Shroyer, K R; Kitatani, K; Obeid, L M; Hannun, Y A

2017-11-23

The protein p38 mitogen-activated protein kinase (MAPK) delta isoform (p38δ) is a poorly studied member of the MAPK family. Data analysis from The Cancer Genome Atlas database revealed that p38δ is highly expressed in all types of human breast cancers. Using a human breast cancer tissue array, we confirmed elevation in cancer tissue. The breast cancer mouse model, MMTV-PyMT (PyMT), developed breast tumors with lung metastasis; however, mice deleted in p38δ (PyMT/p38δ -/- ) exhibited delayed primary tumor formation and highly reduced lung metastatic burden. At the cellular level, we demonstrate that targeting of p38δ in breast cancer cells, MCF-7 and MDA-MB-231 resulted in a reduced rate of cell proliferation. In addition, cells lacking p38δ also displayed an increased cell-matrix adhesion and reduced cell detachment. This effect on cell adhesion was molecularly supported by the regulation of the focal adhesion kinase by p38δ in the human breast cell lines. These studies define a previously unappreciated role for p38δ in breast cancer development and evolution by regulating tumor growth and altering metastatic properties. This study proposes MAPK p38δ protein as a key factor in breast cancer. Lack of p38δ resulted in reduced primary tumor size and blocked the metastatic potential to the lungs.

Aggregation of Culture Expanded Human Mesenchymal Stem Cells in Microcarrier-based Bioreactor.

PubMed

Yuan, Xuegang; Tsai, Ang-Chen; Farrance, Iain; Rowley, Jon; Ma, Teng

2018-03-15

Three-dimensional aggregation of human mesenchymal stem cells (hMSCs) has been used to enhance their therapeutic properties but current fabrication protocols depend on laboratory methods and are not scalable. In this study, we developed thermal responsive poly(N-isopropylacrylamide) grafted microcarriers (PNIPAM-MCs), which supported expansion and thermal detachment of hMSCs at reduced temperature (23.0 °C). hMSCs were cultured on the PNIPAM-MCs in both spinner flask (SF) and PBS Vertical-Wheel (PBS-VW) bioreactors for expansion. At room temperature, hMSCs were detached as small cell sheets, which subsequently self-assembled into 3D hMSC aggregates in PBS-VW bioreactor and remain as single cells in SF bioreactor owing to different hydrodynamic conditions. hMSC aggregates generated from the bioreactor maintained comparable immunomodulation and cytokine secretion properties compared to the ones made from the AggreWell ® . The results of the current study demonstrate the feasibility of scale-up production of hMSC aggregates in the suspension bioreactor using thermal responsive microcarriers for integrated cell expansion and 3D aggregation in a close bioreactor system and highlight the critical role of hydrodynamics in self-assembly of detached hMSC in suspension.

Actomyosin Pulls to Advance the Nucleus in a Migrating Tissue Cell

PubMed Central

Wu, Jun; Kent, Ian A.; Shekhar, Nandini; Chancellor, T.J.; Mendonca, Agnes; Dickinson, Richard B.; Lele, Tanmay P.

2014-01-01

The cytoskeletal forces involved in translocating the nucleus in a migrating tissue cell remain unresolved. Previous studies have variously implicated actomyosin-generated pushing or pulling forces on the nucleus, as well as pulling by nucleus-bound microtubule motors. We found that the nucleus in an isolated migrating cell can move forward without any trailing-edge detachment. When a new lamellipodium was triggered with photoactivation of Rac1, the nucleus moved toward the new lamellipodium. This forward motion required both nuclear-cytoskeletal linkages and myosin activity. Apical or basal actomyosin bundles were found not to translate with the nucleus. Although microtubules dampen fluctuations in nuclear position, they are not required for forward translocation of the nucleus during cell migration. Trailing-edge detachment and pulling with a microneedle produced motion and deformation of the nucleus suggestive of a mechanical coupling between the nucleus and the trailing edge. Significantly, decoupling the nucleus from the cytoskeleton with KASH overexpression greatly decreased the frequency of trailing-edge detachment. Collectively, these results explain how the nucleus is moved in a crawling fibroblast and raise the possibility that forces could be transmitted from the front to the back of the cell through the nucleus. PMID:24411232

Detachment strength of human osteoblasts cultured on hydroxyapatite with various surface roughness. Contribution of integrin subunits.

PubMed

Kokkinos, Petros A; Koutsoukos, Petros G; Deligianni, Despina D

2012-06-01

Hydroxyapatite (HA) has been widely used as a bone substitute in dental, maxillofacial and orthopaedic surgery and as osteoconductive bone substitute or precoating of pedicle screws and cages in spine surgery. The aim of the present study was to investigate the osteoblastic adhesion strength on HA substrata with different surface topography and biochemistry (pre-adsorption of fibronectin) after blocking of specific integrin subunits with monoclonal antibodies. Stoichiometric HA was prepared by precipitation followed by ageing and characterized by SEM, EDX, powder XRD, Raman spectroscopy, TGA, and specific surface area analysis. Human bone marrow derived osteoblasts were cultured on HA disc-shaped substrata which were sintered and polished resulting in two surface roughness grades. For attachment evaluation, cells were incubated with monoclonal antibodies and seeded for 2 h on the substrata. Cell detachment strength was determined using a rotating disc device. Cell detachment strength was surface roughness, fibronectin preadsorption and intergin subunit sensitive.

Inhibiting Myosin Light Chain Kinase Induces Apoptosis In Vitro and In Vivo

PubMed Central

Fazal, Fabeha; Gu, Lianzhi; Ihnatovych, Ivanna; Han, YooJeong; Hu, WenYang; Antic, Nenad; Carreira, Fernando; Blomquist, James F.; Hope, Thomas J.; Ucker, David S.; de Lanerolle, Primal

2005-01-01

Previous short-term studies have correlated an increase in the phosphorylation of the 20-kDa light chain of myosin II (MLC20) with blebbing in apoptotic cells. We have found that this increase in MLC20 phosphorylation is rapidly followed by MLC20 dephosphorylation when cells are stimulated with various apoptotic agents. MLC20 dephosphorylation is not a consequence of apoptosis because MLC20 dephosphorylation precedes caspase activation when cells are stimulated with a proapoptotic agent or when myosin light chain kinase (MLCK) is inhibited pharmacologically or by microinjecting an inhibitory antibody to MLCK. Moreover, blocking caspase activation increased cell survival when MLCK is inhibited or when cells are treated with tumor necrosis factor alpha. Depolymerizing actin filaments or detaching cells, processes that destabilize the cytoskeleton, or inhibiting myosin ATPase activity also resulted in MLC20 dephosphorylation and cell death. In vivo experiments showed that inhibiting MLCK increased the number of apoptotic cells and retarded the growth of mammary cancer cells in mice. Thus, MLC20 dephosphorylation occurs during physiological cell death and prolonged MLC20 dephosphorylation can trigger apoptosis. PMID:15988034

Modification of HTPB-based polyurethane with temperature-sensitive poly(N-isopropyl acrylamide) for biomaterial usage.

PubMed

Yang, Jen Ming; Yang, Shu Jyuan; Lin, Hao Tzu; Chen, Jan Kan

2007-01-01

Hydroxyl-terminated polybutadiene (HTPB)-based polyurethane with dimethyol propionic acid (DPA) as chain extender was synthesized by solution polymerization. The HTPB-based polyurethane was modified by UV radiation with N-isopropyl acrylamide monomer to get poly(N-isopropyl acrylamide)-modified polyurethane (PUDPANIPAAm). The cohesive energy (E(coh)), molar volume (V), solubility parameter (delta), molecular weight (W(M)), volume per gram (V(g)), and the density (1/V(g)) of PUDPANIPAAm were calculated by group contribution methods. To evaluate the application of PUDPANIPAAm for wound dressing and transplantation of cell sheet, the measurement of water content, water vapor transmission rate, and gas permeation on the PUDPANIPAAm membrane was evaluated. The biocompatibility of these membranes, cell adhesion, and proliferation assay were conducted in the cell culture. The effect of thermosensitivity of poly(N-isopropyl acrylamide) on cell detachment was also evaluated in the primary study. The results showed that these PUDPANIPAAm membranes are thermosensitive. The modification of PU with poly(N-isopropyl acrylamide) reduced the water vapor transmission rate and permeability of gas through PUDPANIPAAm membrane. PUDPANIPAAm membranes could support cell adhesion and growth. Owing to the thermosensitive nature of poly(N-isopropyl acrylamide), the relative cell numbers detached from PUDPANIPAAm membranes were larger than those detached from the polystyrene dish. 2006 Wiley Periodicals, Inc.

A study on sonoporation of cells cultured on a soft collagen gel scaffold

NASA Astrophysics Data System (ADS)

Kudo, Nobuki; Kinoshita, Yuto

2012-11-01

Efficiencies of sonoporation were investigated using four types of monolayer-cell samples: cells cultured directly on a cover slip, cells cultured on a cover slip coated with collagen gel of several μm in thickness, and cells cultured on collagen gel scaffolds of 0.4 and 1.0 mm in thicknesses. Cell samples attached with Levovist microbubbles were irradiated by one shot of a three- or 10,000-cycle ultrasound pulse, and cell detachment and membrane perforation were investigated. Experimental results showed that rates of cell detachment and membrane damage were markedly decreased in the presence of soft gel layer of 0.4 and 1.0 mm in thicknesses under the cells and that these rates were inversely proportional to the thickness of the gel layer. These results indicate that optimum conditions of sonoporation in vitro should be carefully translated into those in vivo.

Characterizing the adhesion of motile and nonmotile Escherichia coli to a glass surface using a parallel-plate flow chamber.

PubMed

McClaine, Jennifer W; Ford, Roseanne M

2002-04-20

A parallel-plate flow chamber was used to measure the attachment and detachment rates of Escherichia coli to a glass surface at various fluid velocities. The effect of flagella on adhesion was investigated by performing experiments with several E. coli strains: AW405 (motile); HCB136 (nonmotile mutant with paralyzed flagella); and HCB137 (nonmotile mutant without flagella). We compared the total attachment rates and the fraction of bacteria retained on the surface to determine how the presence and movement of the flagella influence transport to the surface and adhesion strength in this dynamic system. At the lower fluid velocities, there was no significant difference in the total attachment rates for the three bacterial strains; nonmotile strains settled at a rate that was of the same order of magnitude as the diffusion rate of the motile strain. At the highest fluid velocity, the effect of settling was minimized to better illustrate the importance of motility, and the attachment rates of both nonmotile strains were approximately five times slower than that of the motile bacteria. Thus, different processes controlled the attachment rate depending on the parameter regime in which the experiment was performed. The fractions of motile bacteria retained on the glass surface increased with increasing velocity, whereas the opposite trend was found for the nonmotile strains. This suggests that the rotation of the flagella enables cells to detach from the surface (at the lower fluid velocities) and strengthens adhesion (at higher fluid velocities), whereas nonmotile cells detach as a result of shear. There was no significant difference in the initial attachment rates of the two nonmotile species, which suggests that merely the presence of flagella was not important in this stage of biofilm development. Copyright 2002 Wiley Periodicals, Inc.

Transplantation of reprogrammed embryonic stem cells improves visual function in a mouse model for retinitis pigmentosa.

PubMed

Wang, Nan-Kai; Tosi, Joaquin; Kasanuki, Jennifer Mie; Chou, Chai Lin; Kong, Jian; Parmalee, Nancy; Wert, Katherine J; Allikmets, Rando; Lai, Chi-Chun; Chien, Chung-Liang; Nagasaki, Takayuki; Lin, Chyuan-Sheng; Tsang, Stephen H

2010-04-27

To study whether C57BL/6J-Tyr/J (C2J) mouse embryonic stem (ES) cells can differentiate into retinal pigment epithelial (RPE) cells in vitro and then restore retinal function in a model for retinitis pigmentosa: Rpe65/Rpe65 C57BL6 mice. Yellow fluorescent protein (YFP)-labeled C2J ES cells were induced to differentiate into RPE-like structures on PA6 feeders. RPE-specific markers are expressed from differentiated cells in vitro. After differentiation, ES cell-derived RPE-like cells were transplanted into the subretinal space of postnatal day 5 Rpe65/Rpe65 mice. Live imaging of YFP-labeled C2J ES cells demonstrated survival of the graft. Electroretinograms (ERGs) were performed on transplanted mice to evaluate the functional outcome of transplantation. RPE-like cells derived from ES cells sequentially express multiple RPE-specific markers. After transplantation, YFP-labeled cells can be tracked with live imaging for as long as 7 months. Although more than half of the mice were complicated with retinal detachments or tumor development, one fourth of the mice showed increased electroretinogram responses in the transplanted eyes. Rpe65/Rpe65 mice transplanted with RPE-like cells showed significant visual recovery during a 7-month period, whereas those injected with saline, PA6 feeders, or undifferentiated ES cells showed no rescue. ES cells can differentiate, morphologically, and functionally, into RPE-like cells. Based on these findings, differentiated ES cells have the potential for the development of new therapeutic approaches for RPE-specific diseases such as certain forms of retinitis pigmentosa and macular degeneration. Nevertheless, stringent control of retinal detachment and teratoma development will be necessary before initiation of treatment trials.

ICAM-1 and AMPK regulate cell detachment and apoptosis by N-methyl-N Prime -nitro-N-nitrosoguanidine, a widely spread environmental chemical, in human hormone-refractory prostate cancers

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

Chen, Yi-Cheng; Lu, Pin-Hsuan; Hsu, Jui-Ling

2011-12-15

Poly(ADP-ribose) polymerase-1 (PARP-1), a sensor of DNA damage, plays a crucial role in the regulation of DNA repair. PARP-1 hyperactivation causes DNA damage and cell death. The underlying mechanism is complicated and is through diverse pathways. The understanding of responsible signaling pathways may offer implications for effective therapies. After concentration-response determination of N-Methyl-N Prime -Nitro-N-Nitrosoguanidine (MNNG, a PARP-1 activating agent and an environmental mutagen) in human hormone-refractory prostate cancers, the data showed that concentrations below 5 {mu}M did not change cell survival but cause a time-dependent up-regulation of intracellular adhesion molecule-1 (ICAM-1) in mRNA, total protein and cell surface levels.more » Detection of phosphorylation and degradation of I{kappa}B-{alpha} and nuclear translocation of NF-{kappa}B showed that MNNG induced the activation of NF-{kappa}B that was responsible for the ICAM-1 up-regulation since PDTC (a NF-{kappa}B inhibitor) significantly abolished this effect. However, higher concentrations (e.g., 10 {mu}M) of MNNG induced a 61% detachment of the cells which were apoptosis associated with the activation of AMP-activated protein kinase (AMPK), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK). Further identification showed that both AMPK and JNK other than p38 MAPK functionally contributed to cell death. The remaining 39% attached cells were survival associated with high ICAM-1 expression. In conclusion, the data suggest that NF-{kappa}B-dependent up-regulation of ICAM-1 plays a key role on cell attachment and survival; whereas, activation of AMPK and JNK participates in cytotoxic signaling pathways in detached cells caused by PARP-1 activation. Highlights: Black-Right-Pointing-Pointer Low level of DNA damage helps cell attachment and survival via ICAM-1 upregulation. Black-Right-Pointing-Pointer High level of DNA damage causes AMPK- and JNK-involved cell detachment and death. Black-Right-Pointing-Pointer The study provides an anticancer approach targeting PARP-1 and DNA damage response.« less

Citrate, not phosphate, can dissolve calcium oxalate monohydrate crystals and detach these crystals from renal tubular cells.

PubMed

Chutipongtanate, Somchai; Chaiyarit, Sakdithep; Thongboonkerd, Visith

2012-08-15

Dissolution therapy of calcium oxalate monohydrate (COM) kidney stone disease has not yet been implemented due to a lack of well characterized COM dissolution agents. The present study therefore aimed to identify potential COM crystal dissolution compounds. COM crystals were treated with deionized water (negative control), 5 mM EDTA (positive control), 5 mM sodium citrate, or 5mM sodium phosphate. COM crystal dissolution activities of these compounds were evaluated by phase-contrast and video-assisted microscopic examinations, semi-quantitative analysis of crystal size, number and total mass, and spectrophotometric oxalate-dissolution assay. In addition, effects of these compounds on detachment of COM crystals, which adhered tightly onto renal tubular cell surface, were also investigated. The results showed that citrate, not phosphate, had a significant dissolution effect on COM crystals as demonstrated by significant reduction of crystal size (approximately 37% decrease), crystal number (approximately 53% decrease) and total crystal mass (approximately 72% decrease) compared to blank and negative controls. Spectrophotometric oxalate-dissolution assay successfully confirmed the COM crystal dissolution property of citrate. Moreover, citrate could detach up to 85% of the adherent COM crystals from renal tubular cell surface. These data indicate that citrate is better than phosphate for dissolution and detachment of COM crystals. Copyright © 2012 Elsevier B.V. All rights reserved.

Effects of Different Cell-Detaching Methods on the Viability and Cell Surface Antigen Expression of Synovial Mesenchymal Stem Cells.

PubMed

Tsuji, Kunikazu; Ojima, Miyoko; Otabe, Koji; Horie, Masafumi; Koga, Hideyuki; Sekiya, Ichiro; Muneta, Takeshi

2017-06-09

Flow cytometric analysis of cell surface antigens is a powerful tool for the isolation and characterization of stem cells residing in adult tissues. In contrast to the collection of hematopoietic stem cells, the process of enzymatic digestion is usually necessary to prepare mesenchymal stem cells (MSCs) suspensions, which can influence the expression of cell surface markers. In this study, we examined the effects of various cell-detaching reagents and digestion times on the expression of stem cell-related surface antigens and MSC functions. Human MSCs were detached from dishes using four different reagents: trypsin, TrypLE, collagenase, and a nonenzymatic cell dissociation reagent (C5789; Sigma-Aldrich). Following dissociation reagent incubations ranging from 5 to 120 min, cell surface markers were analyzed by flow cytometry. Trypsin and TrypLE quickly dissociated the cells within 5 min, while collagenase and C5789 required 60 min to obtain maximum cell yields. C5789 significantly decreased cell viability at 120 min. Trypsin treatment significantly reduced CD44+, CD55+, CD73+, CD105+, CD140a+, CD140b+, and CD201+ cell numbers within 30 min. Collagenase treatment reduced CD140a expression by 30 min. In contrast, TrypLE treatment did not affect the expression of any cell surface antigens tested by 30 min. Despite the significant loss of surface antigen expression after 60 min of treatment with trypsin, adverse effects of enzymatic digestion on multipotency of MSCs were limited. Overall, our data indicated that TrypLE is advantageous over other cell dissociation reagents tested for the rapid preparation of viable MSC suspensions.

Probing cytoskeletal pre-stress and nuclear mechanics in endothelial cells with spatiotemporally controlled (de-)adhesion kinetics on micropatterned substrates

PubMed Central

Versaevel, Marie; Riaz, Maryam; Corne, Tobias; Grevesse, Thomas; Lantoine, Joséphine; Mohammed, Danahe; Bruyère, Céline; Alaimo, Laura; De Vos, Winnok H.; Gabriele, Sylvain

2017-01-01

ABSTRACT The mechanical properties of living cells reflect their propensity to migrate and respond to external forces. Both cellular and nuclear stiffnesses are strongly influenced by the rigidity of the extracellular matrix (ECM) through reorganization of the cyto- and nucleoskeletal protein connections. Changes in this architectural continuum affect cell mechanics and underlie many pathological conditions. In this context, an accurate and combined quantification of the mechanical properties of both cells and nuclei can contribute to a better understanding of cellular (dys-)function. To address this challenge, we have established a robust method for probing cellular and nuclear deformation during spreading and detachment from micropatterned substrates. We show that (de-)adhesion kinetics of endothelial cells are modulated by substrate stiffness and rely on the actomyosin network. We combined this approach with measurements of cell stiffness by magnetic tweezers to show that relaxation dynamics can be considered as a reliable parameter of cellular pre-stress in adherent cells. During the adhesion stage, large cellular and nuclear deformations occur over a long time span (>60 min). Conversely, nuclear deformation and condensed chromatin are relaxed in a few seconds after detachment. Finally, our results show that accumulation of farnesylated prelamin leads to modifications of the nuclear viscoelastic properties, as reflected by increased nuclear relaxation times. Our method offers an original and non-intrusive way of simultaneously gauging cellular and nuclear mechanics, which can be extended to high-throughput screens of pathological conditions and potential countermeasures. PMID:27111836

Probing cytoskeletal pre-stress and nuclear mechanics in endothelial cells with spatiotemporally controlled (de-)adhesion kinetics on micropatterned substrates.

PubMed

Versaevel, Marie; Riaz, Maryam; Corne, Tobias; Grevesse, Thomas; Lantoine, Joséphine; Mohammed, Danahe; Bruyère, Céline; Alaimo, Laura; De Vos, Winnok H; Gabriele, Sylvain

2017-01-02

The mechanical properties of living cells reflect their propensity to migrate and respond to external forces. Both cellular and nuclear stiffnesses are strongly influenced by the rigidity of the extracellular matrix (ECM) through reorganization of the cyto- and nucleoskeletal protein connections. Changes in this architectural continuum affect cell mechanics and underlie many pathological conditions. In this context, an accurate and combined quantification of the mechanical properties of both cells and nuclei can contribute to a better understanding of cellular (dys-)function. To address this challenge, we have established a robust method for probing cellular and nuclear deformation during spreading and detachment from micropatterned substrates. We show that (de-)adhesion kinetics of endothelial cells are modulated by substrate stiffness and rely on the actomyosin network. We combined this approach with measurements of cell stiffness by magnetic tweezers to show that relaxation dynamics can be considered as a reliable parameter of cellular pre-stress in adherent cells. During the adhesion stage, large cellular and nuclear deformations occur over a long time span (>60 min). Conversely, nuclear deformation and condensed chromatin are relaxed in a few seconds after detachment. Finally, our results show that accumulation of farnesylated prelamin leads to modifications of the nuclear viscoelastic properties, as reflected by increased nuclear relaxation times. Our method offers an original and non-intrusive way of simultaneously gauging cellular and nuclear mechanics, which can be extended to high-throughput screens of pathological conditions and potential countermeasures.

Safety and efficacy of intravitreal injection of recombinant erythropoietin for protection of photoreceptor cells in a rat model of retinal detachment

PubMed Central

Xie, Z; Chen, F; Wu, X; Zhuang, C; Zhu, J; Wang, J; Ji, H; Wang, Y; Hua, X

2012-01-01

Purpose To elucidate the safety and efficacy of exogenous erythropoietin (EPO) for the protection of photoreceptor cells in a rat model of retinal detachment (RD). Methods Recombinant rat EPO (400 ng) was injected into the vitreous cavity of normal rats to observe the eye manifestations. Retinal function was assessed by flash electroretinograms. Histopathological examination of retinal tissue was performed at 14 days and 2 months after injection, respectively. To investigate the inhibitory effect of EPO on photoreceptor cell apoptosis in RD rats, 100, 200, or 400 ng EPO was injected into the vitreous cavity immediately after RD model establishment. Apoptosis of photoreceptor cells was determined at 3 days after injection. Caspase-3 activation was measured by western blot analysis and immunofluorescence, respectively, and the level of Bcl-XL expression was analyzed by western blot. Results Intravitreal injection of EPO 400 ng into normal rats had no significant impact on retinal function, morphology, or structure. Apoptosis of retinal photoreceptor cells apparently increased after RD and was significantly reduced following EPO treatment. The thickness of the outer nuclear layer in the RD+400 ng group was significantly thicker than that in other experimental RD groups both at 14 days and at 2 months after RD (P<0.05). Western blot and immunofluorescence analyses showed decreased caspase-3 activation and increased Bcl-XL expression following EPO treatment. Conclusion Intravitreal injection of EPO 400 ng is safe, and EPO may suppress caspase-3 activation and enhance Bcl-XL expression, resulting in inhibition of apoptosis and protection of photoreceptor cells. PMID:22020175

Visible light controls cell adhesion on a photoswitchable biointerface.

PubMed

Ming, Zunzhen; Hua, Xin; Xue, Yuan; Lin, Qiuning; Bao, Chunyan; Zhu, Linyong

2018-05-04

Bioactive surfaces with specific interactions with cells have been greatly interested due to their potential applications in biosensors and tissue engineering. Herein, we fabricated a dopamine contained photoswitch molecule (compound 1) which could form self-assembled monolayer (SAM) on substrates. The SAM showed a good photoswitch ability and manifested excellent fatigue resistance, which displayed its potential application as a biologically friendly surface coating. Contact angle analysis and cell experiments exhibited that the SAM surface was hydrophobic before irradiation which favored cell adhesion, while, it turned hydrophilic and induced cell unfouling or detachment after light irradiation. The uses of visible light stimulation (λ ex  = 530 nm) and the reversible regulation on cell adhesion and detachment should open up new avenues for bioacitve surfaces in biomedical applications. Copyright © 2018 Elsevier B.V. All rights reserved.

Are core self-evaluations a suitable moderator in stressor-detachment relationships? A study among managers' perceived job demands, detachment and strain reactions.

PubMed

Hentrich, Stephan; Zimber, Andreas; Sosnowsky-Waschek, Nadia; Gregersen, Sabine; Petermann, Franz

2018-01-01

The relationships among job demands, personality factors, recovery and psychological health receive increasing attention but are not well understoodOBJECTIVE:Therefore, the present study tests moderating effects among a sample of managers as proposed by the stressor-detachment model. We aimed to determine whether core self-evaluations (CSE) had an influence on the correlations between detachment and strain reactions (depressive symptoms, irritation, exhaustion) and between job demands and detachment. Further, we tested whether detachment attenuates the positive relation between job demands and strain reactions. A convenience sample of managers in three German settings (N = 282) participated in the cross-sectional study. Results based on hierarchical regression analysis showed that high CSE significantly weakened the negative relationship between detachment and depressive symptoms in this sample. However, CSE did not moderate the negative relationship between job demands and detachment. Moreover, results revealed that detachment moderated the positive relation between job demands and exhaustion. The authors tested whether CSE was able to moderate the relationship between job demands, psychological detachment and different stress reactions. Although we found a significant interaction effect, CSE may be too distal to moderate all respective associations.

Psychological detachment from work during non-work time: linear or curvilinear relations with mental health and work engagement?

PubMed

Shimazu, Akihito; Matsudaira, Ko; Jonge, Jan DE; Tosaka, Naoya; Watanabe, Kazuhiro; Takahashi, Masaya

2016-06-10

This study examined whether a higher level of psychological detachment during non-work time is associated with better employee mental health (Hypothesis 1), and examined whether psychological detachment has a curvilinear relation (inverted U-shaped pattern) with work engagement (Hypothesis 2). A large cross-sectional Internet survey was conducted among registered monitors of an Internet survey company in Japan. The questionnaire included scales for psychological detachment, employee mental health, and work engagement as well as for job characteristics and demographic variables as potential confounders. The hypothesized model was tested with moderated structural equation modeling techniques among 2,234 respondents working in the tertiary industries with regular employment. Results showed that psychological detachment had curvilinear relations with mental health as well as with work engagement. Mental health improved when psychological detachment increased from a low to higher levels but did not benefit any further from extremely high levels of psychological detachment. Work engagement showed the highest level at an intermediate level of detachment (inverted U-shaped pattern). Although high psychological detachment may enhance employee mental health, moderate levels of psychological detachment are most beneficial for his or her work engagement.

Psychological detachment from work during non-work time: linear or curvilinear relations with mental health and work engagement?

PubMed Central

SHIMAZU, Akihito; MATSUDAIRA, Ko; DE JONGE, Jan; TOSAKA, Naoya; WATANABE, Kazuhiro; TAKAHASHI, Masaya

2016-01-01

This study examined whether a higher level of psychological detachment during non-work time is associated with better employee mental health (Hypothesis 1), and examined whether psychological detachment has a curvilinear relation (inverted U-shaped pattern) with work engagement (Hypothesis 2). A large cross-sectional Internet survey was conducted among registered monitors of an Internet survey company in Japan. The questionnaire included scales for psychological detachment, employee mental health, and work engagement as well as for job characteristics and demographic variables as potential confounders. The hypothesized model was tested with moderated structural equation modeling techniques among 2,234 respondents working in the tertiary industries with regular employment. Results showed that psychological detachment had curvilinear relations with mental health as well as with work engagement. Mental health improved when psychological detachment increased from a low to higher levels but did not benefit any further from extremely high levels of psychological detachment. Work engagement showed the highest level at an intermediate level of detachment (inverted U-shaped pattern). Although high psychological detachment may enhance employee mental health, moderate levels of psychological detachment are most beneficial for his or her work engagement. PMID:26829972

Species-dependent hydrodynamics of flagellum-tethered bacteria in early biofilm development.

PubMed

Bennett, Rachel R; Lee, Calvin K; De Anda, Jaime; Nealson, Kenneth H; Yildiz, Fitnat H; O'Toole, George A; Wong, Gerard C L; Golestanian, Ramin

2016-02-01

Monotrichous bacteria on surfaces exhibit complex spinning movements. Such spinning motility is often a part of the surface detachment launch sequence of these cells. To understand the impact of spinning motility on bacterial surface interactions, we develop a hydrodynamic model of a surface-bound bacterium, which reproduces behaviours that we observe in Pseudomonas aeruginosa, Shewanella oneidensis and Vibrio cholerae, and provides a detailed dictionary for connecting observed spinning behaviour to bacteria-surface interactions. Our findings indicate that the fraction of the flagellar filament adhered to the surface, the rotation torque of this appendage, the flexibility of the flagellar hook and the shape of the bacterial cell dictate the likelihood that a microbe will detach and the optimum orientation that it should have during detachment. These findings are important for understanding species-specific reversible attachment, the key transition event between the planktonic and biofilm lifestyle for motile, rod-shaped organisms. © 2016 The Author(s).

Models of dynamic extraction of lipid tethers from cell membranes.

PubMed

Nowak, Sarah A; Chou, Tom

2010-05-07

When a ligand that is bound to an integral membrane receptor is pulled, the membrane and the underlying cytoskeleton can deform before either the membrane delaminates from the cytoskeleton or the ligand detaches from the receptor. If the membrane delaminates from the cytoskeleton, it may be further extruded and form a membrane tether. We develop a phenomenological model for this process by assuming that deformations obey Hooke's law up to a critical force at which the cell membrane locally detaches from the cytoskeleton and a membrane tether forms. We compute the probability of tether formation and show that tethers can be extruded only within an intermediate range of force loading rates and pulling velocities. The mean tether length that arises at the moment of ligand detachment is computed as are the force loading rates and pulling velocities that yield the longest tethers.

CD44S-hyaluronan interactions protect cells resulting from EMT against anoikis

PubMed Central

Cieply, Benjamin; Koontz, Colton; Frisch, Steven M.

2016-01-01

The detachment of normal epithelial cells from matrix triggers an apoptotic response known as anoikis, during homeostatic turnover. Metastatic tumor cells evade anoikis, by mechanisms that are only partly characterized. In particular, the epithelial–mesenchymal transition (EMT) in a subset of invasive tumor cells confers anoikis-resistance. In some cases, EMT up-regulates the cancer stem cell marker CD44S and the enzyme hyaluronic acid synthase-2 (HAS2). CD44S is the major receptor for hyaluronan in the extracellular matrix. Herein, we demonstrate that CD44S, unlike the CD44E isoform expressed in normal epithelial cells, contributes to the protection against anoikis. This protection requires the interaction of CD44S with hyaluronan (HA). CD44S–HA interaction is proposed to play an important role in tumor metastasis through enhanced cell survival under detached conditions. PMID:25937513

Gene Transcription Profile of the Detached Retina (An AOS Thesis)

PubMed Central

Zacks, David N.

2009-01-01

Purpose: Separation of the neurosensory retina from the retinal pigment epithelium (RPE) yields many morphologic and functional consequences, including death of the photoreceptor cells, Müller cell hypertrophy, and inner retinal rewiring. Many of these changes are due to the separation-induced activation of specific genes. In this work, we define the gene transcription profile within the retina as a function of time after detachment. We also define the early activation of kinases that might be responsible for the detachment-induced changes in gene transcription. Methods: Separation of the retina from the RPE was induced in Brown-Norway rats by the injection of 1% hyaluronic acid into the subretinal space. Retinas were harvested at 1, 7, and 28 days after separation. Gene transcription profiles for each time point were determined using the Affymetrix Rat 230A gene microarray chip. Transcription levels in detached retinas were compared to those of nondetached retinas with the BRB-ArrayTools Version 3.6.0 using a random variance analysis of variance (ANOVA) model. Confirmation of the significant transcriptional changes for a subset of the genes was performed using microfluidic quantitative real-time polymerase chain reaction (qRT-PCR) assays. Kinase activation was explored using Western blot analysis to look for early phosphorylation of any of the 3 main families of mitogen-activated protein kinases (MAPK): the p38 family, the Janus kinase family, and the p42/p44 family. Results: Retinas separated from the RPE showed extensive alterations in their gene transcription profile. Many of these changes were initiated as early as 1 day after separation, with significant increases by 7 days. ANOVA analysis defined 144 genes that had significantly altered transcription levels as a function of time after separation when setting a false discovery rate at ≤0.1. Confirmatory RT-PCR was performed on 51 of these 144 genes. Differential transcription detected on the microarray chip was confirmed by qRT-PCR for all 51 genes. Western blot analysis showed that the p42/p44 family of MAPK was phosphorylated within 2 hours of retinal-RPE separation. This phosphorylation was detachment-induced and could be inhibited by specific inhibitors of MAPK phosphorylation. Conclusions: Separation of the retina from the RPE induces significant alteration in the gene transcription profile within the retina. These profiles are not static, but change as a function of time after detachment. These gene transcription changes are preceded by the activation of the p42/p44 family of MAPK. This altered transcription may serve as the basis for many of the morphologic, biochemical, and functional changes seen within the detached retina. PMID:20126507

Burst of reactive oxygen species in pedicel-mediated fruit abscission after carbohydrate supply was cut off in longan (Dimocarpus longan)

PubMed Central

Yang, Ziqin; Zhong, Xiumei; Fan, Yan; Wang, Huicong; Li, Jianguo; Huang, Xuming

2015-01-01

Cutting off carbohydrate supply to longan (Dimocarpus longan Lour.) fruit by girdling and defoliation or by detachment induced 100% abscission within a few days. We used these treatments to study the involvement of reactive oxygen species (ROS) in fruit abscission. Girdling plus defoliation decreased sugar concentrations in the fruit and pedicel and depleted starch grains in the chloroplasts in the cells of abscission zone. Prior to the occurrence of intensive fruit abscission, there was a burst in ROS in the pedicel, which peaked at 1 day after treatment (DAT), when H2O2 in the abscission zone was found to be chiefly located along the plasma membrane (PM). H2O2 was found exclusively in the cell walls 2 DAT, almost disappeared 3 DAT, and reappeared in the mitochondria and cell walls 4 DAT. Signs of cell death such as cytoplasm breakdown were apparent from 3 DAT. The burst of ROS coincided with a sharp increase in the activity of PM-bound NADPH oxidase in the pedicel. At the same time, activities of antioxidant enzymes including superoxide dismutase (SOD), catalase, and peroxidase (POD) were all increased by the treatment and maintained higher than those in the control. Accompanying the reduction in H2O2 abundance, there was a sharp decrease in PM-bound NADPH oxidase activity after 1 DAT in the treated fruit. H2O2 scavenger dimethylthiourea (DMTU, 1 g L–1) significantly inhibited fruit abscission in detached fruit clusters and suppressed the increase in cellulase activity in the abscission zone. These results suggest that fruit abscission induced by carbohydrate stress is mediated by ROS. Roles of ROS in regulating fruit abscission were discussed in relation to its subcellular distribution. PMID:26074931

[Analysis of hydrodynamics parameters of runoff erosion and sediment-yielding on unpaved road].

PubMed

Huang, Peng-Fei; Wang, Wen-Long; Luo, Ting; Wang, Zhen; Wang, Zheng-Li; Li, Ren

2013-02-01

By the method of field runoff washout experiment, a simulation study was conducted on the relationships between the soil detachment rate and the hydrodynamic parameters on unpaved road, and the related quantitative formulas were established. Under the conditions of different flow discharges and road gradients, the averaged soil detachment rate increased with increasing flow discharge and road gradient, and the relationships between them could be described by a power function. As compared with road gradient, flow discharge had greater effects on the soil detachment rate. The soil detachment rate had a power relation with water flow velocity and runoff kinetic energy, and the runoff kinetic energy was of importance to the soil detachment rate. The soil detachment rate was linearly correlated with the unit runoff kinetic energy. The averaged soil erodibility was 0.120 g m-1.J-F-1, and the averaged critical unit runoff kinetic energy was 2.875 g.m-1.J-1. Flow discharge, road gradient, and unit runoff kinetic energy could be used to accurately describe the soil erosion process and calculate the soil erosion rate on unpaved road.

Selective reactivity of monochloramine with extracellular matrix components affects the disinfection of biofilm and detached clusters.

PubMed

Xue, Zheng; Lee, Woo Hyoung; Coburn, Kimberly M; Seo, Youngwoo

2014-04-01

The efficiency of monochloramine disinfection was dependent on the quantity and composition of extracellular polymeric substances (EPS) in biofilms, as monochloramine has a selective reactivity with proteins over polysaccharides. Biofilms with protein-based (Pseudomonas putida) and polysaccharide based EPS (Pseudomonas aeruginosa), as well as biofilms with varied amount of polysaccharide EPS (wild-type and mutant P. aeruginosa), were compared. The different reactivity of EPS components with monochloramine influenced disinfectant penetration, biofilm inactivation, as well as the viability of detached clusters. Monochloramine transport profiling measured by a chloramine-sensitive microelectrode revealed a broader diffusion boundary layer between bulk and biofilm surface in the P. putida biofilm compared to those of P. aeruginosa biofilms. The reaction with proteins in P. putida EPS multiplied both the time and the monochloramine mass required to achieve a full biofilm penetration. Cell viability in biofilms was also spatially influenced by monochloramine diffusion and reaction within biofilms, showing a lower survival in the surface section and a higher persistence in the middle section of the P. putida biofilm compared to the P. aeruginosa biofilms. While polysaccharide EPS promoted biofilm cell viability by obstructing monochloramine reactive sites on bacterial cells, protein EPS hindered monochloramine penetration by reacting with monochloramine and reduced its concentration within biofilms. Furthermore, the persistence of bacterial cells detached from biofilm (over 70% for P. putida and ∼40% for polysaccharide producing P. aeruginosa) suggested that currently recommended monochloramine residual levels may underestimate the risk of water quality deterioration caused by biofilm detachment.

The effect of hypergravity on the lens, cornea and tail regeneration in Urodela

NASA Astrophysics Data System (ADS)

Grigoryan, E. N.; Dvorochkin, N.; Poplinskaya, V. A.; Yousuf, R.; Radugina, E. A.; Almeida, E. A.

2017-09-01

In previous experiments onboard Russian Bion/Foton satellites it was found that exposure to microgravity causes changes in eye lens regeneration of Urodela. The changes included higher rate of regeneration, increased cell proliferation in lens anlage, and synchronization of lens restoration. Similar changes were observed regarding tail regeneration. Recently, investigations were performed to find out whether exposure to hypergravity could also alter lens, cornea and tail regeneration in the newt P. waltl. Nine days prior to exposure the left lens was surgically removed through corneal incision and distal 1/3 of the tail was amputated, thus initiating regeneration. The experimental animals were allowed to recover for 9 days at 1 g and then exposed to 2 g for 12 days in an 8 ft diameter centrifuge at NASA Ames Research Center. The experimental animals were divided into 1 g controls, 2 g centrifugation animals, basal controls, and aquarium controls. Lens and corneal regeneration appeared to be inhibited in 2 g group compared to 1 g animals. In all 1 g controls, lens regeneration reached stages VII-IX in a synchronous fashion and corneal regeneration was nearly complete. In the 2 g newts, neural retinal detachment from the pigmented epithelium was seen in most operated eyes. It was also observed in the non-operated (right) eyes of the animals exposed to 2 g. The level of retinal detachment varied and could have been caused by hypergravity-induced high intraocular pressure. Regeneration (when it could be assessed) proceeded asynchronously, reaching stages from II to IX. Corneal restoration was also noticeably delayed and corneal morphology changed. Cell proliferation was measured using BrdU; the results were not comparable to the 1 g data because of retinal detachment. Previous investigations demonstrated that lens regeneration was controlled by the neural retina; therefore, lower regeneration rate at 2 g was, at least in part, associated with retinal detachment. FGF2 release by the neural retina and FGFR2 expression in the iris and other tissues could accelerate lens proliferation whereas its delay could be caused by retinal detachment, which may explain compromised regeneration at 2 g. Hypergravity (both 1 g and 2 g) increased tissue growth compared to aquarium control (as measured by regenerate volume) and altered the shape of tail regenerates - they became curved downwards. The experimental results emphasize the important and versatile role gravity plays in tissue regeneration. They also suggest that, when considering hypergravity as a countermeasure that can be used in future space missions, its potential impact on the eye should not be ignored.

Protein Breakdown and Formation of Protease in Attached and Detached Cotyledons of Phaseolus vulgaris L.

PubMed

Yomo, H; Srinivasan, K

1973-12-01

In contrast to earlier reported results of similar experiments in peas, in which almost no increase in protease activity occurred in incubated detached cotyledons, we report here an increase in protease activity in both attached and detached bean cotyledons. Detached bean cotyledons showed continually increasing protease activity up to the 12th day, while that in attached cotyledons declined after 6 days. The free amino acid level in detached cotyledons reached a maximum at the 11th day; protease formation leveled off after 50% of the original seed protein was digested. These data suggest that high free amino acid levels may inhibit protease formation.The activity of partially purified protease in aqueous extracts was enhanced by 10 mm 2-mercaptoethanol or cysteine, indicating a sulfhydryl requirement for activation. Protease formation in detached cotyledons was inhibited 30% by 10 mug/ml cycloheximide and 50% by 100 mum abscisic acid. In contrast, alpha-amylase formation was inhibited 90% by 10 mug/ml cycloheximide and 95% by 20 mum abscisic acid. The cycloheximide data suggest that only a part of the protease, but all of the alpha-amylase, is synthesized de novo; the similar pattern of inhibition by abscisic acid emphasizes the concept that protease may exist in two forms.

Protein Breakdown and Formation of Protease in Attached and Detached Cotyledons of Phaseolus vulgaris L. 12

PubMed Central

Yomo, Harugoro; Srinivasan, Komala

1973-01-01

In contrast to earlier reported results of similar experiments in peas, in which almost no increase in protease activity occurred in incubated detached cotyledons, we report here an increase in protease activity in both attached and detached bean cotyledons. Detached bean cotyledons showed continually increasing protease activity up to the 12th day, while that in attached cotyledons declined after 6 days. The free amino acid level in detached cotyledons reached a maximum at the 11th day; protease formation leveled off after 50% of the original seed protein was digested. These data suggest that high free amino acid levels may inhibit protease formation. The activity of partially purified protease in aqueous extracts was enhanced by 10 mm 2-mercaptoethanol or cysteine, indicating a sulfhydryl requirement for activation. Protease formation in detached cotyledons was inhibited 30% by 10 μg/ml cycloheximide and 50% by 100 μm abscisic acid. In contrast, α-amylase formation was inhibited 90% by 10 μg/ml cycloheximide and 95% by 20 μm abscisic acid. The cycloheximide data suggest that only a part of the protease, but all of the α-amylase, is synthesized de novo; the similar pattern of inhibition by abscisic acid emphasizes the concept that protease may exist in two forms. PMID:16658628

Mitochondrial clearance by the STK38 kinase supports oncogenic Ras-induced cell transformation

PubMed Central

Bettoun, Audrey; Surdez, Didier; Vallerand, David; Gundogdu, Ramazan; Sharif, Ahmad A.D.; Gomez, Marta; Cascone, Ilaria; Meunier, Brigitte; White, Michael A.; Codogno, Patrice; Parrini, Maria Carla; Camonis, Jacques H.; Hergovich, Alexander

2016-01-01

Oncogenic Ras signalling occurs frequently in many human cancers. However, no effective targeted therapies are currently available to treat patients suffering from Ras-driven tumours. Therefore, it is imperative to identify downstream effectors of Ras signalling that potentially represent promising new therapeutic options. Particularly, considering that autophagy inhibition can impair the survival of Ras-transformed cells in tissue culture and mouse models, an understanding of factors regulating the balance between autophagy and apoptosis in Ras-transformed human cells is needed. Here, we report critical roles of the STK38 protein kinase in oncogenic Ras transformation. STK38 knockdown impaired anoikis resistance, anchorage-independent soft agar growth, and in vivo xenograft growth of Ras-transformed human cells. Mechanistically, STK38 supports Ras-driven transformation through promoting detachment-induced autophagy. Even more importantly, upon cell detachment STK38 is required to sustain the removal of damaged mitochondria by mitophagy, a selective autophagic process, to prevent excessive mitochondrial reactive oxygen species production that can negatively affect cancer cell survival. Significantly, knockdown of PINK1 or Parkin, two positive regulators of mitophagy, also impaired anoikis resistance and anchorage-independent growth of Ras-transformed human cells, while knockdown of USP30, a negative regulator of PINK1/Parkin-mediated mitophagy, restored anchorage-independent growth of STK38-depleted Ras-transformed human cells. Therefore, our findings collectively reveal novel molecular players that determine whether Ras-transformed human cells die or survive upon cell detachment, which potentially could be exploited for the development of novel strategies to target Ras-transformed cells. PMID:27283898

Transient Proliferation of Proanthocyanidin-Accumulating Cells on the Epidermal Apex Contributes to Highly Aluminum-Resistant Root Elongation in Camphor Tree1[W

PubMed Central

Osawa, Hiroki; Endo, Izuki; Hara, Yukari; Matsushima, Yuki; Tange, Takeshi

2011-01-01

Aluminum (Al) is a harmful element that rapidly inhibits the elongation of plant roots in acidic soils. The release of organic anions explains Al resistance in annual crops, but the mechanisms that are responsible for superior Al resistance in some woody plants remain unclear. We examined cell properties at the surface layer of the root apex in the camphor tree (Cinnamomum camphora) to understand its high Al resistance mechanism. Exposure to 500 μm Al for 8 d, more than 20-fold higher concentration and longer duration than what soybean (Glycine max) can tolerate, only reduced root elongation in the camphor tree to 64% of the control despite the slight induction of citrate release. In addition, Al content in the root apices was maintained at low levels. Histochemical profiling revealed that proanthocyanidin (PA)-accumulating cells were present at the adjacent outer layer of epidermis cells at the root apex, having distinctive zones for cell division and the early phase of cell expansion. Then the PA cells were gradually detached off the root, leaving thin debris behind, and the root surface was replaced with the elongating epidermis cells at the 3- to 4-mm region behind the tip. Al did not affect the proliferation of PA cells or epidermis cells, except for the delay in the start of expansion and the accelerated detachment of the former. In soybean roots, the innermost lateral root cap cells were absent in both PA accumulation and active cell division and failed to protect the epidermal cell expansion at 25 μm Al. These results suggest that transient proliferation and detachment of PA cells may facilitate the expansion of epidermis cells away from Al during root elongation in camphor tree. PMID:21045123

Transient proliferation of proanthocyanidin-accumulating cells on the epidermal apex contributes to highly aluminum-resistant root elongation in camphor tree.

PubMed

Osawa, Hiroki; Endo, Izuki; Hara, Yukari; Matsushima, Yuki; Tange, Takeshi

2011-01-01

Aluminum (Al) is a harmful element that rapidly inhibits the elongation of plant roots in acidic soils. The release of organic anions explains Al resistance in annual crops, but the mechanisms that are responsible for superior Al resistance in some woody plants remain unclear. We examined cell properties at the surface layer of the root apex in the camphor tree (Cinnamomum camphora) to understand its high Al resistance mechanism. Exposure to 500 μm Al for 8 d, more than 20-fold higher concentration and longer duration than what soybean (Glycine max) can tolerate, only reduced root elongation in the camphor tree to 64% of the control despite the slight induction of citrate release. In addition, Al content in the root apices was maintained at low levels. Histochemical profiling revealed that proanthocyanidin (PA)-accumulating cells were present at the adjacent outer layer of epidermis cells at the root apex, having distinctive zones for cell division and the early phase of cell expansion. Then the PA cells were gradually detached off the root, leaving thin debris behind, and the root surface was replaced with the elongating epidermis cells at the 3- to 4-mm region behind the tip. Al did not affect the proliferation of PA cells or epidermis cells, except for the delay in the start of expansion and the accelerated detachment of the former. In soybean roots, the innermost lateral root cap cells were absent in both PA accumulation and active cell division and failed to protect the epidermal cell expansion at 25 μm Al. These results suggest that transient proliferation and detachment of PA cells may facilitate the expansion of epidermis cells away from Al during root elongation in camphor tree.

Ag+ alters cell growth, neurite extension, cardiomyocyte beating, and fertilized egg constriction.

PubMed

Conrad, A H; Tramp, C R; Long, C J; Wells, D C; Paulsen, A Q; Conrad, G W

1999-11-01

The Russian Space Agency uses electrochemically generated silver ions (Ag+) to purify drinking water for their space station, Mir, and their portion of the International Space Station. U.S. EPA guidelines allow 10.6 micromol x L(-1) Ag+ in human drinking water for up to 10 d. Studies correlate Ag+ exposure with tissue dysfunction in humans, rats, and mice, and with altered ion transport, skeletal muscle contraction, and embryonic cell constriction in other animal cells. Ag+ effects on cell shape change-related functions have not been assessed. Immortalized embryonic human intestinal epithelial cells, freshly explanted embryonic avian nerve cells and cardiomyocytes, and marine fertilized eggs were grown in vitro in medium containing AgNO3. Intestinal cells detach from the substratum and viable cell number decreases by 5-6 d at 5 micromol x L(-1) AgNO3, and faster at higher concentrations. Microtubules appear unaltered in adherent cells. Detached cells are nonviable. Neurite outgrowth and glial cell migration from dorsal root ganglia are inhibited by 3 d at 15 micromol x L(-1) AgNO3 or greater. Contractions stop temporarily in most cardiomyocytes by 5 min at 5 micromol x L(-1) AgNO3 or more, but some cardiomyocytes beat 3 times faster than normal at 7.5-20 micromol x L(-1) AgNO3. Picomolar Ag+ increases marine egg polar lobe constriction within an hour, even in the absence of microtubules. Ag+ alters animal cell growth and shape changes by a MT-independent mechanism. This is the first report of Ag+ effects on vertebrate neurite outgrowth, glial cell migration, or cardiomyocyte beat rate.

Prevention of retinal detachment in Stickler syndrome: the Cambridge prophylactic cryotherapy protocol.

PubMed

Fincham, Gregory S; Pasea, Laura; Carroll, Christopher; McNinch, Annie M; Poulson, Arabella V; Richards, Allan J; Scott, John D; Snead, Martin P

2014-08-01

The Stickler syndromes are the most common causes of inherited and childhood retinal detachment; however, no consensus exists regarding the effectiveness of prophylactic intervention. We evaluate the long-term safety and efficacy of the Cambridge prophylactic cryotherapy protocol, a standardized retinal prophylactic treatment developed to prevent retinal detachment arising from giant retinal tears in type 1 Stickler syndrome. Retrospective comparative case series. Four hundred eighty seven patients with type 1 Stickler syndrome. Time to retinal detachment was compared between patients who received bilateral prophylaxis and untreated controls, with and without individual patient matching. Patients receiving unilateral prophylaxis (after fellow eye retinal detachment) were similarly compared with an appropriate control subgroup. Individual patient matching ensured equal age and follow-up between groups and that an appropriate control (who had not suffered a retinal detachment before the age at which their individually matched treatment patient underwent prophylactic treatment) was selected. Matching was blinded to outcome events. Individual patient matching protocols purposely weighted bias against the effectiveness of treatment. All treatment side effects are reported. Time to retinal detachment and side effects occurring after prophylactic treatment. The bilateral control group (n = 194) had a 7.4-fold increased risk of retinal detachment compared to the bilateral prophylaxis group (n = 229) (hazard ratio [HR], 7.40; 95% confidence interval [CI], 4.53-12.08; P<0.001); the matched bilateral control group (n = 165) had a 5.0-fold increased risk compared to the matched bilateral prophylaxis group (n = 165) (HR, 4.97; 95% CI, 2.82-8.78; P<0.001). The unilateral control group (n = 104) had a 10.3-fold increased risk of retinal detachment compared to the unilateral prophylaxis group (n = 64) (HR, 10.29; 95% CI, 4.96-21.36; P<0.001); the matched unilateral control group (n = 39) had a 8.4-fold increased risk compared to the matched unilateral prophylaxis group (n = 39) (HR, 8.36; 95% CI, 3.24-21.57; P<0.001). No significant long-term side effects occurred. In the largest global cohort of type 1 Stickler syndrome patients published, all analyses indicate that the Cambridge prophylactic cryotherapy protocol is safe and markedly reduces the risk of retinal detachment. Copyright © 2014 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.

Retinal detachment following endophthalmitis.

PubMed

Nelsen, P T; Marcus, D A; Bovino, J A

1985-08-01

Fifty-five consecutive patients with a clinical diagnosis of bacterial endophthalmitis were reviewed. All patients were treated with systemic, periocular, topical, and intravitreal antibiotics. In addition, 33 of the patients underwent a pars plana vitrectomy. Nine retinal detachments occurred within six months of initial diagnosis. The higher frequency of retinal detachment in the vitrectomy group (21%) as compared to those patients managed without vitrectomy (9%) may be explained by a combination of surgical complications and the increased severity of endophthalmitis in the vitrectomy group. The two patients who developed retinal detachment during vitrectomy surgery rapidly progressed to no light perception. Conversely, the repair of retinal detachments diagnosed postoperatively had a good prognosis.

Mammalian Auditory Hair Cell Bundle Stiffness Affects Frequency Tuning by Increasing Coupling along the Length of the Cochlea.

PubMed

Dewey, James B; Xia, Anping; Müller, Ulrich; Belyantseva, Inna A; Applegate, Brian E; Oghalai, John S

2018-06-05

The stereociliary bundles of cochlear hair cells convert mechanical vibrations into the electrical signals required for auditory sensation. While the stiffness of the bundles strongly influences mechanotransduction, its influence on the vibratory response of the cochlear partition is unclear. To assess this, we measured cochlear vibrations in mutant mice with reduced bundle stiffness or with a tectorial membrane (TM) that is detached from the sensory epithelium. We found that reducing bundle stiffness decreased the high-frequency extent and sharpened the tuning of vibratory responses obtained postmortem. Detaching the TM further reduced the high-frequency extent of the vibrations but also lowered the partition's resonant frequency. Together, these results demonstrate that the bundle's stiffness and attachment to the TM contribute to passive longitudinal coupling in the cochlea. We conclude that the stereociliary bundles and TM interact to facilitate passive-wave propagation to more apical locations, possibly enhancing active-wave amplification in vivo. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

Novel Approach to Measuring the Droplet Detachment Force from Fibers.

PubMed

Amrei, M M; Venkateshan, D G; D'Souza, N; Atulasimha, J; Tafreshi, H Vahedi

2016-12-20

Determining the force required to detach a droplet from a fiber or from an assembly of fibers is of great importance to many applications. A novel technique is developed in this work to measure this force experimentally by using ferrofluid droplets in a magnetic field. Unlike previous methods reported in the literature, our technique does not require air flow or a mechanical object to detach the droplet from the fiber(s); therefore, it simplifies the experiment and also allows one to study the capillarity of the droplet-fiber system in a more isolated environment. In this article, we investigated the effects of the relative angle between intersecting fibers on the force required to detach a droplet from the fibers in the in-plane or out-of-plane direction. The in-plane and through-plane detachment forces were also predicted via numerical simulation and compared with the experimental results. Good agreement was observed between the numerical and experimental results. It was found that the relative angle between intersecting fibers has no significant effect on the detachment force in the out-of-plane direction. However, the detachment force in the in-plane direction depends strongly on the relative angle between the fibers, and it increases as this angle increases.

Dynamics of Single Hydrogen Bubbles at a Platinum Microelectrode.

PubMed

Yang, Xuegeng; Karnbach, Franziska; Uhlemann, Margitta; Odenbach, Stefan; Eckert, Kerstin

2015-07-28

Bubble dynamics, including the formation, growth, and detachment, of single H2 bubbles was studied at a platinum microelectrode during the electrolysis of 1 M H2SO4 electrolyte. The bubbles were visualized through a microscope by a high-speed camera. Electrochemical measurements were conducted in parallel to measure the transient current. The periodic current oscillations, resulting from the periodic formation and detachment of single bubbles, allow the bubble lifetime and size to be predicted from the transient current. A comparison of the bubble volume calculated from the current and from the recorded bubble image shows a gas evolution efficiency increasing continuously with the growth of the bubble until it reaches 100%. Two different substrates, glass and epoxy, were used to embed the Pt wire. While nearly no difference was found with respect to the growth law for the bubble radius, the contact angle differs strongly for the two types of cell. Data provided for the contact point evolution further complete the image of single hydrogen bubble growth. Finally, the velocity field driven by the detached bubble was measured by means of PIV, and the effects of the convection on the subsequent bubble were evaluated.

Chemical and physical effects on the adhesion, maturation, and survival of monocytes, macrophages, and foreign body giant cells

NASA Astrophysics Data System (ADS)

Collier, Terry Odell, III

Injury caused by biomedical device implantation initiates inflammatory and wound healing responses. Cells migrate to the site of injury to degrade bacteria and toxins, create new vasculature, and form new and repair injured tissue. Blood-proteins rapidly adsorb onto the implanted material surface and express adhesive ligands which mediate cell adhesion on the material surface. Monocyte-derived macrophages and multi-nucleated foreign body giant cells adhere to the surface and degrade the surface of the material. Due to the role of macrophage and foreign body giant cell on material biocompatibility and biostability, the effects of surface chemistry, surface topography and specific proteins on the maturation and survival of monocytes, macrophages and foreign body giant cells has been investigated. Novel molecularly designed materials were used to elucidate the dynamic interactions which occur between inflammatory cells, proteins and surfaces. The effect of protein and protein adhesion was investigated using adhesive protein depleted serum conditions on RGD-modified and silane modified surfaces. The effects of surface chemistry were investigated using temperature responsive surfaces of poly (N-isopropylacrylamide) and micropatterned surfaces of N-(2 aminoethyl)-3-aminopropyltrimethoxysilane regions on an interpenetrating polymer network of polyacrylamide and poly(ethylene glycol). The physical effects were investigated using polyimide scaffold materials and polyurethane materials with surface modifying end groups. The depletion of immunoglobulin G caused decreased levels of macrophage adhesion, foreign body giant cell formation and increased levels of apoptosis. The temporal nature of macrophage adhesion was observed with changing effectiveness of adherent cell detachment with time, which correlated to increased expression of beta1 integrin receptors on detached macrophages with time. The limited ability of the micropatterned surface, polyimide scaffold and surface modified polyurethane materials to control macrophage adhesion indicates the complexity of macrophage adhesion and protein adsorption onto a surface. These studies have indicated components and adhesive mechanisms which can be utilized to create materials with enhanced resistance to macrophage adhesion and/or degradative abilities.

Heat shock protein 27 regulates human prostate cancer cell motility and metastatic progression

PubMed Central

Voll, Eric A; Ogden, Irene M; Pavese, Janet M; Huang, XiaoKe; Xu, Li; Jovanovic, Borko D; Bergan, Raymond C

2014-01-01

Prostate cancer (PCa) is the most common form of cancer in American men. Mortality from PCa is caused by the movement of cancer cells from the primary organ to form metastatic tumors at distant sites. Heat shock protein 27 (HSP27) is known to increase human PCa cell invasion and its overexpression is associated with metastatic disease. The role of HSP27 in driving PCa cell movement from the prostate to distant metastatic sites is unknown. Increased HSP27 expression increased metastasis as well as primary tumor mass. In vitro studies further examined the mechanism of HSP27-induced metastatic behavior. HSP27 did not affect cell detachment, adhesion, or migration, but did increase cell invasion. Cell invasion was dependent upon matrix metalloproteinase 2 (MMP-2), whose expression was increased by HSP27. In vivo, HSP27 induced commensurate changes in MMP-2 expression in tumors. These findings demonstrate that HSP27 drives metastatic spread of cancer cells from the prostate to distant sites, does so across a continuum of expression levels, and identifies HSP27-driven increases in MMP-2 expression as functionally relevant. These findings add to prior studies demonstrating that HSP27 increases PCa cell motility, growth and survival. Together, they demonstrate that HSP27 plays an important role in PCa progression. PMID:24798191

A Taz1- and Microtubule-Dependent Regulatory Relationship between Telomere and Centromere Positions in Bouquet Formation Secures Proper Meiotic Divisions

PubMed Central

Katsumata, Kazuhiro; Hirayasu, Ami; Miyoshi, Junpei; Nishi, Eriko; Ichikawa, Kento; Tateho, Kazuki; Wakuda, Airi; Matsuhara, Hirotada; Yamamoto, Ayumu

2016-01-01

During meiotic prophase, telomeres cluster, forming the bouquet chromosome arrangement, and facilitate homologous chromosome pairing. In fission yeast, bouquet formation requires switching of telomere and centromere positions. Centromeres are located at the spindle pole body (SPB) during mitotic interphase, and upon entering meiosis, telomeres cluster at the SPB, followed by centromere detachment from the SPB. Telomere clustering depends on the formation of the microtubule-organizing center at telomeres by the linker of nucleoskeleton and cytoskeleton complex (LINC), while centromere detachment depends on disassembly of kinetochores, which induces meiotic centromere formation. However, how the switching of telomere and centromere positions occurs during bouquet formation is not fully understood. Here, we show that, when impaired telomere interaction with the LINC or microtubule disruption inhibited telomere clustering, kinetochore disassembly-dependent centromere detachment and accompanying meiotic centromere formation were also inhibited. Efficient centromere detachment required telomere clustering-dependent SPB recruitment of a conserved telomere component, Taz1, and microtubules. Furthermore, when artificial SPB recruitment of Taz1 induced centromere detachment in telomere clustering-defective cells, spindle formation was impaired. Thus, detachment of centromeres from the SPB without telomere clustering causes spindle impairment. These findings establish novel regulatory mechanisms, which prevent concurrent detachment of telomeres and centromeres from the SPB during bouquet formation and secure proper meiotic divisions. PMID:27611693

Fabrication of thermo-responsive PNIPAAm-g-ETFE for cell culture dishes by pre-irradiation grafting

NASA Astrophysics Data System (ADS)

Yamahara, Yumi; Nagasawa, Naotsugu; Taguchi, Mitsumasa; Oshima, Akihiro; Washio, Masakazu

2018-01-01

Thermo-responsive templates for the cell cultivation based on Poly(tetrafluoroethylene-co-ethylene) (ETFE) were fabricated by pre-irradiation grafting of N-isoproplyacrylamide (NIPAAm) monomer by electron beam (EB) irradiation under nitrogen gas atmosphere at room temperature, and their characteristic properties were studied. The detachment of cultured HeLa cells from fabricated thermo-responsive templates were attempted. Furthermore, the reaction mechanism is proposed using ESR spectroscopy and FT-IR spectroscopy. It is confirmed that the cultured HeLa cells were detached from fabricated thermo-responsive templates at 20 °C. Water contact angle analysis indicated that obtained templates had thermo-response around 30 °C. It is suggested that the grafted polymer chains would mainly react with peroxy radicals (-CF2-CF(OO･)-) on tetrafluoroethylene unit in ETFE.

Transconjunctival drainage of serous and hemorrhagic choroidal detachment.

PubMed

Rezende, Flávio A; Kickinger, Mônica C; Li, Gisèle; Prado, Renata F; Regis, Luiz Gustavo T

2012-02-01

To describe a novel surgical technique for drainage of bullous serous and hemorrhagic choroidal detachments. A prospective, consecutive case series of 6 eyes with serous and/or hemorrhagic choroidal detachments secondary to intraocular surgery was documented to evaluate the feasibility of using the 25-gauge and 20-gauge transconjunctival trocar/cannula systems to drain choroidal detachments. Two eyes had expulsive hemorrhagic choroidal detachments and 4 eyes had serous choroidal detachments after glaucoma surgeries. A 25-gauge infusion line was placed in the anterior chamber. A 20-gauge (in eyes with hemorrhagic choroidal detachments) or a 25-gauge (in eyes with serous detachments) trocar/cannula system was inserted into the suprachoroidal space 7.0 mm from limbus. After drainage, the cannulas were removed and no sutures were placed. Pars plana vitrectomy was performed only in eyes with concomitant pathology that demanded the additional procedure. The primary outcome measure was presence of choroidal detachment at 1 week, 2 weeks, and 1 month postoperatively. Secondary outcome measures were visual acuity at 6 months and intraocular pressure at 1 week and 1, 3, and 6 months postoperatively. Drainage of hemorrhagic choroidal detachments resulted in resolution of the detachments by 1 month postoperatively. In eyes with serous detachments, resolution was achieved by 1 week postdrainage. In both groups, intraocular pressure increased to at least 10 mmHg by postoperative Week 1. The visual acuity improved in all eyes. No complications related to the transconjunctival technique were noted. Transconjunctival drainage of serous and hemorrhagic choroidal detachments seems to be a feasible and simple surgical option with minimal scleral and conjunctival damage. Pars plana vitrectomy may not be necessary when draining choroidal detachments in this manner.

Residual Gas Effects on Detached Solidification in Microgravity

NASA Technical Reports Server (NTRS)

Wilcox, William R.; Regel, Liya L.; Ramakrishnan; Kota, Arun; Anand, Gaurav

2004-01-01

Our long term goal has been to make detached solidification reproducible, which requires a full understanding of the mechanisms underlying it. Our Moving Meniscus Model of steady-state detachment predicts that it depends strongly on the surface tension of the melt and the advancing contact angle with the ampoule wall. Thus, the objective of the current project was to determine the influence of residual gases on the surface tension and contact angle of molten semiconductors on typical ampoule materials. Our focus was on the influence of oxygen on indium antimonide on clean silica ("quartz"). The research was performed by three chemical engineering graduate students, the third of whom will complete his research in the summer of 2005. Originally, we had planned to use a sealed silica cell containing a zirconia electrochemical element to control the oxygen partial pressure. However, zirconia requires an operating temperature above the 530 C melting point of InSb and is difficult to form a gas-tight seal with silica. Thus, we decided instead to flow an oxygen-containing gas through the cell. A special apparatus was designed, built and perfected. A piece of InSb was placed on a horizontal silica plate in a quartz cell. High purity argon, helium or hydrogen-containing gas is passed continuously through the cell while the oxygen concentration in the effluent gas is measured. The shape of the resulting drop was used to determine contact angle and surface tension of Ga-doped and high purity InSb. Oxygen appeared to decrease the contact angle, and definitely did not increase it. The following section gives the background for the research. Section 2 summarizes the results obtained on Ga-doped InSb with relatively high oxygen concentrations. Section 3 describes recent improvements made to the apparatus and methods of analysis. Section 4 gives recent results for high-purity InSb at low oxygen concentrations. Final results will be obtained only this summer (2005). Each section has its own references.

Accelerated podocyte detachment and progressive podocyte loss from glomeruli with age in Alport Syndrome.

PubMed

Ding, Fangrui; Wickman, Larysa; Wang, Su Q; Zhang, Yanqin; Wang, Fang; Afshinnia, Farsad; Hodgin, Jeffrey; Ding, Jie; Wiggins, Roger C

2017-12-01

Podocyte depletion is a common mechanism driving progression in glomerular diseases. Alport Syndrome glomerulopathy, caused by defective α3α4α5 (IV) collagen heterotrimer production by podocytes, is associated with an increased rate of podocyte detachment detectable in urine and reduced glomerular podocyte number suggesting that defective podocyte adherence to the glomerular basement membrane might play a role in driving progression. Here a genetically phenotyped Alport Syndrome cohort of 95 individuals [urine study] and 41 archived biopsies [biopsy study] were used to test this hypothesis. Podocyte detachment rate (measured by podocin mRNA in urine pellets expressed either per creatinine or 24-hour excretion) was significantly increased 11-fold above control, and prior to a detectably increased proteinuria or microalbuminuria. In parallel, Alport Syndrome glomeruli lose an average 26 podocytes per year versus control glomeruli that lose 2.3 podocytes per year, an 11-fold difference corresponding to the increased urine podocyte detachment rate. Podocyte number per glomerulus in Alport Syndrome biopsies is projected to be normal at birth (558/glomerulus) but accelerated podocyte loss was projected to cause end-stage kidney disease by about 22 years. Biopsy data from two independent cohorts showed a similar estimated glomerular podocyte loss rate comparable to the measured 11-fold increase in podocyte detachment rate. Reduction in podocyte number and density in biopsies correlated with proteinuria, glomerulosclerosis, and reduced renal function. Thus, the podocyte detachment rate appears to be increased from birth in Alport Syndrome, drives the progression process, and could potentially help predict time to end-stage kidney disease and response to treatment. Copyright © 2017 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.

A novel platform for minimally invasive delivery of cellular therapy as a thin layer across the subretina for treatment of retinal degeneration

NASA Astrophysics Data System (ADS)

Rotenstreich, Ygal; Tzameret, Adi; Kalish, Sapir E.; Belkin, Michael; Meir, Amilia; Treves, Avraham J.; Nagler, Arnon; Sher, Ifat

2015-03-01

Incurable retinal degenerations affect millions worldwide. Stem cell transplantation rescued visual functions in animal models of retinal degeneration. In those studies cells were transplanted in subretinal "blebs", limited number of cells could be injected and photoreceptor rescue was restricted to areas in proximity to the injection sites. We developed a minimally-invasive surgical platform for drug and cell delivery in a thin layer across the subretina and extravascular spaces of the choroid. The novel system is comprised of a syringe with a blunt-tipped needle and an adjustable separator. Human bone marrow mesenchymal stem cells (hBM-MSCs) were transplanted in eyes of RCS rats and NZW rabbits through a longitudinal triangular scleral incision. No immunosuppressants were used. Retinal function was determined by electroretinogram analysis and retinal structure was determined by histological analysis and OCT. Transplanted cells were identified as a thin layer across the subretina and extravascular spaces of the choroid. In RCS rats, cell transplantation delayed photoreceptor degeneration across the entire retina and significantly enhanced retinal functions. No retinal detachment or choroidal hemorrhages were observed in rabbits following transplantation. This novel platform opens a new avenue for drug and cell delivery, placing the transplanted cells in close proximity to the damaged RPE and retina as a thin layer, across the subretina and thereby slowing down cell death and photoreceptor degeneration, without retinal detachment or choroidal hemorrhage. This new transplantation system may increase the therapeutic effect of other cell-based therapies and therapeutic agents. This study is expected to directly lead to phase I/II clinical trials for autologous hBM-MSCs transplantation in retinal degeneration patients.

Thermo-responsive methylcellulose hydrogels as temporary substrate for cell sheet biofabrication.

PubMed

Altomare, Lina; Cochis, Andrea; Carletta, Andrea; Rimondini, Lia; Farè, Silvia

2016-05-01

Methylcellulose (MC), a water-soluble polymer derived from cellulose, was investigated as a possible temporary substrate having thermo-responsive properties favorable for cell culturing. MC-based hydrogels were prepared by a dispersion technique, mixing MC powder (2, 4, 6, 8, 10, 12 % w/v) with selected salts (sodium sulphate, Na2SO4), sodium phosphate, calcium chloride, or phosphate buffered saline, to evaluate the influence of different compositions on the thermo-responsive behavior. The inversion test was used to determine the gelation temperatures of the different hydrogel compositions; thermo-mechanical properties and thermo-reversibility of the MC hydrogels were investigated by rheological analysis. Gelation temperatures and rheological behavior depended on the MC concentration and type and concentration of salt used in hydrogel preparation. In vitro cytotoxicity tests, performed using L929 mouse fibroblasts, showed no toxic release from all the tested hydrogels. Among the investigated compositions, the hydrogel composed of 8 % w/v MC with 0.05 M Na2SO4 had a thermo-reversibility temperature at 37 °C. For that reason, this formulation was thus considered to verify the possibility of inducing in vitro spontaneous detachment of cells previously seeded on the hydrogel surface. A continuous cell layer (cell sheet) was allowed to grow and then detached from the hydrogel surface without the use of enzymes, thanks to the thermo-responsive behavior of the MC hydrogel. Immunofluorescence observation confirmed that the detached cell sheet was composed of closely interacting cells.

Importance of anisotropy in detachment rates for force production and cargo transport by a team of motor proteins.

PubMed

Takshak, Anjneya; Kunwar, Ambarish

2016-05-01

Many cellular processes are driven by collective forces generated by a team consisting of multiple molecular motor proteins. One aspect that has received less attention is the detachment rate of molecular motors under mechanical force/load. While detachment rate of kinesin motors measured under backward force increases rapidly for forces beyond stall-force; this scenario is just reversed for non-yeast dynein motors where detachment rate from microtubule decreases, exhibiting a catch-bond type behavior. It has been shown recently that yeast dynein responds anisotropically to applied load, i.e. detachment rates are different under forward and backward pulling. Here, we use computational modeling to show that these anisotropic detachment rates might help yeast dynein motors to improve their collective force generation in the absence of catch-bond behavior. We further show that the travel distance of cargos would be longer if detachment rates are anisotropic. Our results suggest that anisotropic detachment rates could be an alternative strategy for motors to improve the transport properties and force production by the team. © 2016 The Protein Society.

Acute tissue reactions, inner segment pathology, and effects of the antioxidant α1-microglobulin in an in vitro model of retinal detachment.

PubMed

Ghosh, Fredrik; Åkerström, Bo; Bergwik, Jesper; Abdshill, Hodan; Gefors, Lina; Taylor, Linnéa

2018-04-18

The purpose of this study was to explore acute tissue reactions, ultrastructural photoreceptor morphology with emphasis on inner segments, and the effect of antioxidant treatment in an in vitro model of rhegmatogenous retinal detachment (RRD). A previously described method of RRD simulation was used with adult retinal porcine explants kept free-floating in culture medium with or without treatment with the radical scavenger α 1 -microglobulin (A1M). Explants were examined at 5 time points from 1 to 24 h using transmission electron microscopy as well as quantitative real-time PCR (RT-PCR) to quantify gene expression of the cell stress marker heat shock protein 70 (Hsp70) and oxidative stress marker heme oxygenase (HO-1). The culture medium level of the cell damage marker lactate dehydrogenase (LDH) and oxidative stress DNA damage marker 8-Oxo-2'-deoxyguanosine (8-OHdG) was also assessed at each time point. We found that the levels of Hsp70 and LDH rapidly increased in both groups, and at 3 and 6 h, Hsp70 was significantly higher in A1M treated retinas. At 24 h, Hsp70 and LDH, as well as 8-OHdG were significantly lower compared with controls, whereas the tissue level of HO-1 was significantly higher. Progressive ultrastructural photoreceptor changes were seen in untreated control explants from 1 h and onwards including outer segment shortening and loss, disruption of organelles within the inner segments and loss of perikarya in the outer nuclear layer. Inner segment pathology was more rapid and extensive in rods compared with in cones. In A1M treated counterparts, damage to rod inner segment mitochondria was significantly higher after 1 h of culture, but after this time, no statistical difference was found. At 24 h, cone inner segment mitochondrial disruption was significantly higher in control retinas and the number of surviving perikarya lower. From our results, we conclude that retinal explants elicit acute cell stress reactions when placed in culture without physical support simulating a detached retina floating in the vitreous space. Photoreceptors rapidly display degenerative changes including extensive damage to inner segment mitochondria indicating loss of energy transduction as an early key event. A1M increases initial mitochondrial stress in the rods, however, subsequent pathology is attenuated by the treatment, highlighting the dynamics of protective as well as disruptive oxidative stress reactions in the detached retina. Copyright © 2018 Elsevier Ltd. All rights reserved.

Dynamics of Bovine Sperm Interaction with Epithelium Differ Between Oviductal Isthmus and Ampulla1

PubMed Central

Ardon, Florencia; Markello, Ross D.; Hu, Lian; Deutsch, Zarah I.; Tung, Chih-Kuan; Wu, Mingming; Suarez, Susan S.

2016-01-01

In mammals, many sperm that reach the oviduct are held in a reservoir by binding to epithelium. To leave the reservoir, sperm detach from the epithelium; however, they may bind and detach again as they ascend into the ampulla toward oocytes. In order to elucidate the nature of binding interactions along the oviduct, we compared the effects of bursts of strong fluid flow (as would be caused by oviductal contractions), heparin, and hyperactivation on detachment of bovine sperm bound in vitro to epithelium on intact folds of isthmic and ampullar mucosa. Intact folds of oviductal mucosa were used to represent the strong attachments of epithelial cells to each other and to underlying connective tissue that exist in vivo. Effects of heparin on binding were tested because heparin binds to the Binder of SPerm (BSP) proteins that attach sperm to oviductal epithelium. Sperm bound by their heads to beating cilia on both isthmic and ampullar epithelia and could not be detached by strong bursts of fluid flow. Addition of heparin immediately detached sperm from isthmic epithelium but not ampullar epithelium. Addition of 4-aminopyridine immediately stimulated hyperactivation of sperm but did not detach them from isthmic or ampullar epithelium unless added with heparin. These observations indicate that the nature of binding of sperm to ampullar epithelium differs from that of binding to isthmic epithelium; specifically, sperm bound to isthmic epithelium can be detached by heparin alone, while sperm bound to ampullar epithelium requires both heparin and hyperactivation to detach from the epithelium. PMID:27605344

Dynamics of Bovine Sperm Interaction with Epithelium Differ Between Oviductal Isthmus and Ampulla.

PubMed

Ardon, Florencia; Markello, Ross D; Hu, Lian; Deutsch, Zarah I; Tung, Chih-Kuan; Wu, Mingming; Suarez, Susan S

2016-10-01

In mammals, many sperm that reach the oviduct are held in a reservoir by binding to epithelium. To leave the reservoir, sperm detach from the epithelium; however, they may bind and detach again as they ascend into the ampulla toward oocytes. In order to elucidate the nature of binding interactions along the oviduct, we compared the effects of bursts of strong fluid flow (as would be caused by oviductal contractions), heparin, and hyperactivation on detachment of bovine sperm bound in vitro to epithelium on intact folds of isthmic and ampullar mucosa. Intact folds of oviductal mucosa were used to represent the strong attachments of epithelial cells to each other and to underlying connective tissue that exist in vivo. Effects of heparin on binding were tested because heparin binds to the Binder of SPerm (BSP) proteins that attach sperm to oviductal epithelium. Sperm bound by their heads to beating cilia on both isthmic and ampullar epithelia and could not be detached by strong bursts of fluid flow. Addition of heparin immediately detached sperm from isthmic epithelium but not ampullar epithelium. Addition of 4-aminopyridine immediately stimulated hyperactivation of sperm but did not detach them from isthmic or ampullar epithelium unless added with heparin. These observations indicate that the nature of binding of sperm to ampullar epithelium differs from that of binding to isthmic epithelium; specifically, sperm bound to isthmic epithelium can be detached by heparin alone, while sperm bound to ampullar epithelium requires both heparin and hyperactivation to detach from the epithelium. © 2016 by the Society for the Study of Reproduction, Inc.

Modalities to enumerate circulating tumor cells in the bloodstream for cancer prognosis and to monitor the response to the therapy.

PubMed

Romano, G

2017-09-01

Certain malignant cells may detach from the primary tumor and enter the vascular system, forming so-called circulating tumor cells (CTCs). Clusters of malignant cells associated with other cell types can also be observed in the peripheral blood of oncological patients. Such cell clusters are termed circulating tumor microemboli (CTM). The isolation and quantification of CTCs and/or CTM from blood samples allow for an accurate prognosis of the clinical course of the disease and to monitor the response to therapy. Current protocols rely on epithelial markers for the isolation of CTCs and/or CTM from hematopoietic cells. However, epithelial markers may be silenced during the progression of the epithelial-mesenchymal transition, which regulates the detachment and migration of malignant cells from the primary tumor. This review summarizes the achievements and challenges of various modalities for the isolation, enrichment, analysis and enumeration of CTCs and/or CTM, in order to assess the advancement of the disease and the response to therapy.

Improved Crystal Quality By Detached Solidification in Microgravity

NASA Technical Reports Server (NTRS)

Regel, Liya L.; Wilcox, William R.; Wang, Yaz-Hen; Wang, Jian-Bin

2003-01-01

Many microgravity directional solidification experiments yielded ingots with portions that grew without contacting the ampoule wall, leading to greatly improved crystallographic perfection. Our long term goals have been: (1) To develop a complete understanding of all of the phenomena of detached solidification.; (2) To make it possible to achieve detached solidification reproducibly; (3) To increase crystallographic perfection through detached solidification. We have three major achievements to report here: (1) We obtained a new material balance solution for the Moving Meniscus Model of detached solidification. This solution greatly clarifies the physics as well as the roles of the parameters in the system; (2) We achieved detached solidification of InSb growing on earth in BN-coated ampoules; (3) We performed an extensive series of experiments on freezing water that showed how to form multiple gas bubbles or tubes on the ampoule wall. However, these did not propagate around the wall and lead to fully detached solidification unless the ampoule wall was extremely rough and non-wetted.

Clearance of apoptotic photoreceptors: elimination of apoptotic debris into the subretinal space and macrophage-mediated phagocytosis via phosphatidylserine receptor and integrin alphavbeta3.

PubMed

Hisatomi, Toshio; Sakamoto, Taiji; Sonoda, Koh-Hei; Tsutsumi, Chikako; Qiao, Hong; Enaida, Hiroshi; Yamanaka, Ichiro; Kubota, Toshiaki; Ishibashi, Tatsuro; Kura, Shinobu; Susin, Santos A; Kroemer, Guido

2003-06-01

The effective phagocytotic clearance of apoptotic debris is fundamental to the maintenance of neural tissues during apoptosis. Retinal photoreceptors undergo apoptosis after retinal detachment. Although their induction phase of apoptosis has been well discussed, their phagocytotic process remains quite unclear. We herein demonstrate that apoptotic photoreceptors are selectively eliminated from their physiological localization, the outer nuclear layer, to the subretinal space, and then phagocytosed by monocyte-derived macrophages. This could be shown by an ultrastructural and immunophenotypic analysis. Moreover, in chimera mice expressing transgenic green fluorescent protein in bone marrow-derived cells, the local infiltration of macrophages could be detected after retinal detachment-induced photoreceptor apoptosis. The local injection of an antibody blocking the phosphatidylserine receptor (PSR) or a peptide (GRGDSP)-blocking integrin alphavbeta3 revealed that phagocytotic clearance involves the PSR as well as integrin alphavbeta3 in vivo. Importantly, the level of blockade obtained with these reagents was different. Although anti-PSR increased the frequency of apoptotic cells that fail to bind to macrophages, GRGDSP prevented the engulfment (but not the recognition) of apoptotic photoreceptor cells by macrophages. To our knowledge, this is the first report describing the mechanisms through which apoptotic photoreceptors are selectively eliminated via a directional process in the subretinal space.

Clearance of Apoptotic Photoreceptors

PubMed Central

Hisatomi, Toshio; Sakamoto, Taiji; Sonoda, Koh-hei; Tsutsumi, Chikako; Qiao, Hong; Enaida, Hiroshi; Yamanaka, Ichiro; Kubota, Toshiaki; Ishibashi, Tatsuro; Kura, Shinobu; Susin, Santos A.; Kroemer, Guido

2003-01-01

The effective phagocytotic clearance of apoptotic debris is fundamental to the maintenance of neural tissues during apoptosis. Retinal photoreceptors undergo apoptosis after retinal detachment. Although their induction phase of apoptosis has been well discussed, their phagocytotic process remains quite unclear. We herein demonstrate that apoptotic photoreceptors are selectively eliminated from their physiological localization, the outer nuclear layer, to the subretinal space, and then phagocytosed by monocyte-derived macrophages. This could be shown by an ultrastructural and immunophenotypic analysis. Moreover, in chimera mice expressing transgenic green fluorescent protein in bone marrow-derived cells, the local infiltration of macrophages could be detected after retinal detachment-induced photoreceptor apoptosis. The local injection of an antibody blocking the phosphatidylserine receptor (PSR) or a peptide (GRGDSP)-blocking integrin αvβ3 revealed that phagocytotic clearance involves the PSR as well as integrin αvβ3 in vivo. Importantly, the level of blockade obtained with these reagents was different. Although anti-PSR increased the frequency of apoptotic cells that fail to bind to macrophages, GRGDSP prevented the engulfment (but not the recognition) of apoptotic photoreceptor cells by macrophages. To our knowledge, this is the first report describing the mechanisms through which apoptotic photoreceptors are selectively eliminated via a directional process in the subretinal space. PMID:12759244

Rat supraspinatus muscle atrophy after tendon detachment.

PubMed

Barton, Elisabeth R; Gimbel, Jonathan A; Williams, Gerald R; Soslowsky, Louis J

2005-03-01

Rotator cuff tears are one of the most common tendon disorders found in the healthy population. Tendon tears not only affect the biomechanical properties of the tendon, but can also lead to debilitation of the muscles attached to the damaged tendons. The changes that occur in the muscle after tendon detachment are not well understood. A rat rotator cuff model was utilized to determine the time course of changes that occur in the supraspinatus muscle after tendon detachment. It was hypothesized that the lack of load on the supraspinatus muscle would cause a significant decrease in muscle mass and a conversion of muscle fiber properties toward those of fast fiber types. Tendons were detached at the insertion on the humerus without repair. Muscle mass, morphology and fiber properties were measured at one, two, four, eight, and 16 weeks after detachment. Tendon detachment resulted in a rapid loss of muscle mass, an increase in the proportion of fast muscle fibers, and an increase in the fibrotic content of the muscle bed, concomitant with the appearance of adhesions of the tendon to surrounding surfaces. At 16 weeks post-detachment, muscle mass and the fiber properties in the deep muscle layers returned to normal levels. However, the fiber shifts observed in the superficial layers persisted throughout the experiment. These results suggest that load returned to the muscle via adhesions to surrounding surfaces, which may be sufficient to reverse changes in muscle mass.

Electron detachment of the hydrogen-bonded amino acid side-chain guanine complexes

NASA Astrophysics Data System (ADS)

Wang, Jing; Gu, Jiande; Leszczynski, Jerzy

2007-07-01

The photoelectron spectra of the hydrogen-bonded amino acid side-chain-guanine complexes has been studied at the partial third order (P3) self-energy approximation of the electron propagator theory. The correlation between the vertical electron detachment energy and the charge distributions on the guanine moiety reveals that the vertical electron detachment energy (VDE) increases as the positive charge distribution on the guanine increases. The low VDE values determined for the negatively charged complexes of the guanine-side-chain-group of Asp/Glu suggest that the influence of the H-bonded anionic groups on the VDE of guanine could be more important than that of the anionic backbone structure. The even lower vertical electron detachment energy for guanine is thus can be expected in the H-bonded protein-DNA systems.

Detachment-induced E-cadherin expression promotes 3D tumor spheroid formation but inhibits tumor formation and metastasis of lung cancer cells.

PubMed

Powan, Phattrakorn; Luanpitpong, Sudjit; He, Xiaoqing; Rojanasakul, Yon; Chanvorachote, Pithi

2017-11-01

The epithelial-to-mesenchymal transition is proposed to be a key mechanism responsible for metastasis-related deaths. Similarly, cancer stem cells (CSCs) have been proposed to be a key driver of tumor metastasis. However, the link between the two events and their control mechanisms is unclear. We used a three-dimensional (3D) tumor spheroid assay and other CSC-indicating assays to investigate the role of E-cadherin in CSC regulation and its association to epithelial-to-mesenchymal transition in lung cancer cells. Ectopic overexpression and knockdown of E-cadherin were found to promote and retard, respectively, the formation of tumor spheroids in vitro but had opposite effects on tumor formation and metastasis in vivo in a xenograft mouse model. We explored the discrepancy between the in vitro and in vivo results and demonstrated, for the first time, that E-cadherin is required as a component of a major survival pathway under detachment conditions. Downregulation of E-cadherin increased the stemness of lung cancer cells but had an adverse effect on their survival, particularly on non-CSCs. Such downregulation also promoted anoikis resistance and invasiveness of lung cancer cells. These results suggest that anoikis assay could be used as an alternative method for in vitro assessment of CSCs that involves dysregulated adhesion proteins. Our data also suggest that agents that restore E-cadherin expression may be used as therapeutic agents for metastatic cancers. Copyright © 2017 the American Physiological Society.

CUB domain-containing protein 1 and the epidermal growth factor receptor cooperate to induce cell detachment.

PubMed

Law, Mary E; Ferreira, Renan B; Davis, Bradley J; Higgins, Paul J; Kim, Jae-Sung; Castellano, Ronald K; Chen, Sixue; Luesch, Hendrik; Law, Brian K

2016-08-05

While localized malignancies often respond to available therapies, most disseminated cancers are refractory. Novel approaches, therefore, are needed for the treatment of metastatic disease. CUB domain-containing protein1 (CDCP1) plays an important role in metastasis and drug resistance; the mechanism however, is poorly understood. Breast cancer cell lines were engineered to stably express EGFR, CDCP1 or phosphorylation site mutants of CDCP1. These cell lines were used for immunoblot analysis or affinity purification followed by immunoblot analysis to assess protein phosphorylation and/or protein complex formation with CDCP1. Kinase activity was evaluated using phosphorylation site-specific antibodies and immunoblot analysis in in vitro kinase assays. Protein band excision and mass spectrometry was utilized to further identify proteins complexed with CDCP1 or ΔCDCP1, which is a mimetic of the cleaved form of CDCP1. Cell detachment was assessed using cell counting. This paper reports that CDCP1 forms ternary protein complexes with Src and EGFR, facilitating Src activation and Src-dependent EGFR transactivation. Importantly, we have discovered that a class of compounds termed Disulfide bond Disrupting Agents (DDAs) blocks CDCP1/EGFR/Src ternary complex formation and downstream signaling. CDCP1 and EGFR cooperate to induce detachment of breast cancer cells from the substratum and to disrupt adherens junctions. Analysis of CDCP1-containing complexes using proteomics techniques reveals that CDCP1 associates with several proteins involved in cell adhesion, including adherens junction and desmosomal cadherins, and cytoskeletal elements. Together, these results suggest that CDCP1 may facilitate loss of adhesion by promoting activation of EGFR and Src at sites of cell-cell and cell-substratum contact.

Design of a novel bioreactor and application in vascular tissue engineering

NASA Astrophysics Data System (ADS)

Zhang, Zhi-Xiong; Xi, Ting-Fei; Wang, Ying-Jun; Chen, Xiao-Song; Zhang, Jian; Wang, Chun-Ren; Gu, Yong-Quan; Chen, Liang; Li, Jian-Xin; Chen, Bing

2008-11-01

Endothelial cells (ECs) detachment under high shear stress at the early period of transplantation resulted in thrombosis and occlusion. To solve this problem, we developed a novel bioreactor. The bioreactor mimicked the formation of pulsatile flow in physiological conditions. Human umbilical vein ECs were seeded onto the lumen of living tissue conduits grown within dog peritoneal cavity. The shear stress generated by the bioreactor was increased step by step from 1.5 ± 0.8 dyn/cm 2 to 5.3 ± 2.4 dyn/cm 2, and was applied to ECs after static culture for 2 days. The results showed that completely confluent monolayer ECs were elongated, and were oriented parallel to the flow direction. The bioreactor could provide good environment for formation of endothelium. Stepwise increase shear stress could strengthen cell-cell and cell-extracellular matrix. The flow conditions of the bioreactor play a key role to determine the quality of the ECs lining.

Data of continuous harvest of stem cells via partial detachment from thermoresponsive nanobrush surfaces

PubMed Central

Yeh, Chin-Chen; Muduli, Saradaprasan; Peng, I-Chia; Lu, Yi-Tung; Ling, Qing-Dong; Alarfaj, Abdullah A.; Munusamy, Murugan A.; Kumar, S. Suresh; Murugan, Kadarkarai; Chen, Da-Chung; Lee, Hsin-chung; Chang, Yung; Higuchi, Akon

2016-01-01

This data article contains two figures and one table supporting the research article entitled: “Continuous harvest of stem cells via partial detachment from thermoresponsive nanobrush surface” [1]. The table shows coating conditions of three copolymers, poly(styrene-co-acrylic acid) grafted with oligovitronectin, poly(styrene-co-N-isopropylacrylamide) and poly(styrene-co-polyethylene glycol methacrylate) to prepare thermoresponsive surface. XPS spectra show the nitrogen peak of the polystyrene surface coated with poly(styrene-co-acrylic acid) grafted with oligovitronectin. The surface coating density analyzed from sorption of poly(styrene-co-acrylic acid) grafted with oligovitronectin by UV–vis spectroscopy is also presented. PMID:26909373

Blastocyst production after intracytoplasmic sperm injection with semen from a stallion with testicular degeneration.

PubMed

Roels, K; Smits, K; Ververs, C; Govaere, J; D'Herde, K; Van Soom, A

2018-06-01

In horse breeding, intracytoplasmic sperm injection (ICSI) has gained interest to obtain offspring from subfertile individuals. This paper presents a case report of a stallion with severe testicular degeneration. Semen analysis showed very low motility and 83.5% of detached heads. Histology of a testicular biopsy showed severely decreased spermatogenesis, while transmission electron microscopy of the sperm cells revealed no significant abnormalities. A total of 39 oocytes were fertilized by ICSI with frozen-thawed spermatozoa of this stallion: 25 oocytes with intact spermatozoa and 24 with detached heads. When using intact sperm cells, 8 out of the 25 oocytes cleaved, and 1 developed to the blastocyst stage 9 days after ICSI. None of the oocytes injected with a detached sperm head cleaved. Studies on the paternal influence on ICSI outcome are limited in the horse and further research is needed to define which stallion factors may influence ICSI results. Here, we report the possibility to produce a blastocyst by ICSI of a stallion suffering from testicular degeneration with a poor spermiogram, as long as an intact sperm cell containing a centriole is selected. © 2018 Blackwell Verlag GmbH.

Microbial cell surface characteristics: Elucidating attachment/detachment using hydrophobicity and electrokinetic measurements

EPA Science Inventory

The surface properties of microorganisms play an important role in their behavior within the environment. Electrophoretic mobility and cell surface hydrophobicity of bacterial cells influence their initial interaction with surfaces and mediate their stability within an aqueous su...

TGF-β regulates LARG and GEF-H1 during EMT to affect stiffening response to force and cell invasion

PubMed Central

Osborne, Lukas D.; Li, George Z.; How, Tam; O'Brien, E. Tim; Blobe, Gerard C.; Superfine, Richard; Mythreye, Karthikeyan

2014-01-01

Recent studies implicate a role for cell mechanics in cancer progression. The epithelial-to-mesenchymal transition (EMT) regulates the detachment of cancer cells from the epithelium and facilitates their invasion into stromal tissue. Although classic EMT hallmarks include loss of cell–cell adhesions, morphology changes, and increased invasion capacity, little is known about the associated mechanical changes. Previously, force application on integrins has been shown to initiate cytoskeletal rearrangements that result in increased cell stiffness and a stiffening response. Here we demonstrate that transforming growth factor β (TGF-β)–induced EMT results in decreased stiffness and loss of the normal stiffening response to force applied on integrins. We find that suppression of the RhoA guanine nucleotide exchange factors (GEFs) LARG and GEF-H1 through TGF-β/ALK5–enhanced proteasomal degradation mediates these changes in cell mechanics and affects EMT-associated invasion. Taken together, our results reveal a functional connection between attenuated stiffness and stiffening response and the increased invasion capacity acquired after TGF-β–induced EMT. PMID:25143398

Bilateral patching in retinal detachment: fluid mechanics and retinal "settling".

PubMed

Foster, William J

2011-07-20

When a patient suffers a retinal detachment and surgery is delayed, it is known clinically that bilaterally patching the patient may allow the retina to partially reattach or "settle." Although this procedure has been performed since the 1860s, there is still debate as to how such a maneuver facilitates the reattachment of the retina. Finite element calculations using commercially available analysis software are used to elucidate the influence of reduction in eye movement caused by bilateral patching on the flow of subretinal fluid in a physical model of retinal detachment. It was found that by coupling fluid mechanics with structural mechanics, a physically consistent explanation of increased retinal detachment with eye movements can be found in the case of traction on the retinal hole. Large eye movements increase vitreous traction and detachment forces on the edge of the retinal hole, creating a subretinal vacuum and facilitating increased subretinal fluid. Alternative models, in which intraocular fluid flow is redirected into the subretinal space, are not consistent with these simulations. The results of these simulations explain the physical principles behind bilateral patching and provide insight that can be used clinically. In particular, as is known clinically, bilateral patching may facilitate a decrease in the height of a retinal detachment. The results described here provide a description of a physical mechanism underlying this technique. The findings of this study may aid in deciding whether to bilaterally patch patients and in counseling patients on pre- and postoperative care.

Paediatric retinal detachment: aetiology, characteristics and outcomes.

PubMed

McElnea, Elizabeth; Stephenson, Kirk; Gilmore, Sarah; O'Keefe, Michael; Keegan, David

2018-01-01

To provide contemporary data on the aetiology, clinical features and outcomes of paediatric retinal detachment. A retrospective review of all those under 16y who underwent surgical repair for retinal detachment at a single centre between the years 2008 and 2015 inclusive was performed. In each case the cause of retinal detachment, the type of detachment, the presence or absence of macular involvement, the number and form of reparative surgeries undertaken, and the surgical outcome achieved was recorded. Twenty-eight eyes of 24 patients, 15 (62.5%) of whom were male and 9 (37.5%) of whom were female, their mean age being 11.6y and range 2-16y developed retinal detachment over the eight year period studied. Trauma featured in the development of retinal detachment in 14 (50.0%) cases. Retinal detachment was associated with other ocular and/or systemic conditions in 11 (39.3%) cases. A mean of 3.0 procedures with a range of 1-9 procedures per patient were undertaken in the management of retinal detachment. Complex vitrectomy combined with scleral buckling or complex vitrectomy alone were those most frequently performed. Mean postoperative visual acuity was 1.2 logMAR with range 0.0-3.0 logMAR. In 22 of 26 (84.6%) cases which underwent surgical repair the retina was attached at last follow-up. Aggressive management of paediatric retinal detachment including re-operation increases the likelihood of anatomical success. In cases where the retinal detachment can be repaired by an external approach alone there is a more favourable visual outcome.

SLC12A7 alters adrenocortical carcinoma cell adhesion properties to promote an aggressive invasive behavior.

PubMed

Brown, Taylor C; Murtha, Timothy D; Rubinstein, Jill C; Korah, Reju; Carling, Tobias

2018-06-08

Altered expression of Solute Carrier Family 12 Member 7 (SLC12A7) is implicated to promote malignant behavior in multiple cancer types through an incompletely understood mechanism. Recent studies have shown recurrent gene amplifications and overexpression of SLC12A7 in adrenocortical carcinoma (ACC). The potential mechanistic effect(s) of SLC12A7 amplifications in portending an aggressive behavior in ACC has not been previously studied and is investigated here using two established ACC cell lines, SW-13 and NCI-H295R. SW-13 cells, which express negligible amounts of SLC12A7, were enforced to express SLC12A7 constitutively, while RNAi gene silencing was performed in NCI-H295R cells, which have robust endogenous expression of SLC12A7. In vitro studies tested the outcomes of experimental alterations in SLC12A7 expression on malignant characteristics, including cell viability, growth, colony formation potential, motility, invasive capacity, adhesion and detachment kinetics, and cell membrane organization. Further, potential alterations in transcription regulation downstream to induced SLC12A7 overexpression was explored using targeted transcription factor expression arrays. Enforced SLC12A7 overexpression in SW-13 cells robustly promoted motility and invasive characteristics (p < 0.05) without significantly altering cell viability, growth, or colony formation potential. SLC12A7 overexpression also significantly increased rates of cellular attachment and detachment turnover (p < 0.05), potentially propelled by increased filopodia formation and/or Ezrin interaction. In contrast, RNAi gene silencing of SLC12A7 stymied cell attachment strength as well as migration and invasion capacity in NCI-H295R cells. Transcription factor expression analysis identified multiple signally pathways potentially affected by SLC12A7 overexpression, including osmotic stress, bone morphogenetic protein, and Hippo signaling pathways. Amplification of SLC12A7 observed in ACCs is shown here, in vitro, to exacerbate the malignant behavior of ACC cells by promoting invasive capacities, possibly mediated by alterations in multiple signaling pathways, including the osmotic stress pathway.

Microphysical modeling of Titan's detached haze layer in a 3D GCM

NASA Astrophysics Data System (ADS)

Larson, Erik J. L.; Toon, Owen B.; West, Robert A.; Friedson, A. James

2015-07-01

We use a 3D GCM with coupled aerosol microphysics to investigate the formation and seasonal cycle of the detached haze layer in Titan's upper atmosphere. The base of the detached haze layer is defined by a local minimum in the vertical extinction profile. The detached haze is seen at all latitudes including the south pole as seen in Cassini images from 2005-2012. The layer merges into the winter polar haze at high latitudes where the Hadley circulation carries the particles downward. The hemisphere in which the haze merges with the polar haze varies with season. We find that the base of the detached haze layer occurs where there is a near balance between vertical winds and particle fall velocities. Generally the vertical variation of particle concentration in the detached haze region is simply controlled by sedimentation, so the concentration and the extinction vary roughly in proportion to air density. This variation explains why the upper part of the main haze layer, and the bulk of the detached haze layer follow exponential profiles. However, the shape of the profile is modified in regions where the vertical wind velocity is comparable to the particle fall velocity. Our simulations closely match the period when the base of the detached layer in the tropics is observed to begin its seasonal drop in altitude, and the total range of the altitude drop. However, the simulations have the base of the detached layer about 100 km lower than observed, and the time for the base to descend is slower in the simulations than observed. These differences may point to the model having somewhat lower vertical winds than occur on Titan, or somewhat too large of particle sizes, or some combination of both. Our model is consistent with a dynamical origin for the detached haze rather than a chemical or microphysical one. This balance between the vertical wind and particle fall velocities occurs throughout the summer hemisphere and tropics. The particle concentration gradients that are established in the summer hemisphere are transported to the winter hemisphere by meridional winds from the overturning Hadley cell. Our model is consistent with the disappearance of the detached haze layer in early 2014. Our simulations predict the detached haze and gap will reemerge at its original high altitude between mid 2014 and early 2015.

Inhibition of Late and Early Phases of Cancer Metastasis by the NF-κB Inhibitor DHMEQ Derived from Microbial Bioactive Metabolite Epoxyquinomicin: A Review.

PubMed

Lin, Yinzhi; Ukaji, Tamami; Koide, Naoki; Umezawa, Kazuo

2018-03-03

We previously designed and synthesized dehydroxyepoxyquinomicin (DHMEQ) as an inhibitor of NF-κB based on the structure of microbial secondary metabolite epoxyquinomicin C. DHMEQ showed anti-inflammatory and anticancer activity in various in vivo disease models without toxicity. On the other hand, the process of cancer metastasis consists of cell detachment from the primary tumor, invasion, transportation by blood or lymphatic vessels, invasion, attachment, and formation of secondary tumor. Cell detachment from the primary tumor and subsequent invasion are considered to be early phases of metastasis, while tumor cell attachment to the tissue and secondary tumor formation the late phases. The assay system for the latter phase was set up with intra-portal-vein injection of pancreatic cancer cells. Intraperitoneal administration of DHMEQ was found to inhibit liver metastasis possibly by decreasing the expression of MMP-9 and IL-8. Also, when the pancreatic cancer cells treated with DHMEQ were inoculated into the peritoneal cavity of mice, the metastatic foci formation was inhibited. These results indicate that DHMEQ is likely to inhibit the late phase of metastasis. Meanwhile, we have recently employed three-dimensional (3D) culture of breast cancer cells for the model of early phase metastasis, since the 3D invasion just includes cell detachment and invasion into the matrix. DHMEQ inhibited the 3D invasion of breast cancer cells at 3D-nontoxic concentrations. In this way, DHMEQ was shown to inhibit the late and early phases of metastasis. Thus, DHMEQ is likely to be useful for the suppression of cancer metastasis.

Directing Stem Cell Differentiation via Electrochemical Reversible Switching between Nanotubes and Nanotips of Polypyrrole Array.

PubMed

Wei, Yan; Mo, Xiaoju; Zhang, Pengchao; Li, Yingying; Liao, Jingwen; Li, Yongjun; Zhang, Jinxing; Ning, Chengyun; Wang, Shutao; Deng, Xuliang; Jiang, Lei

2017-06-27

Control of stem cell behaviors at solid biointerfaces is critical for stem-cell-based regeneration and generally achieved by engineering chemical composition, topography, and stiffness. However, the influence of dynamic stimuli at the nanoscale from solid biointerfaces on stem cell fate remains unclear. Herein, we show that electrochemical switching of a polypyrrole (Ppy) array between nanotubes and nanotips can alter surface adhesion, which can strongly influence mechanotransduction activation and guide differentiation of mesenchymal stem cells (MSCs). The Ppy array, prepared via template-free electrochemical polymerization, can be reversibly switched between highly adhesive hydrophobic nanotubes and poorly adhesive hydrophilic nanotips through an electrochemical oxidation/reduction process, resulting in dynamic attachment and detachment to MSCs at the nanoscale. Multicyclic attachment/detachment of the Ppy array to MSCs can activate intracellular mechanotransduction and osteogenic differentiation independent of surface stiffness and chemical induction. This smart surface, permitting transduction of nanoscaled dynamic physical inputs into biological outputs, provides an alternative to classical cell culture substrates for regulating stem cell fate commitment. This study represents a general strategy to explore nanoscaled interactions between stem cells and stimuli-responsive surfaces.

Increased heat dissipation with the X-divertor geometry facilitating detachment onset at lower density in DIII-D

NASA Astrophysics Data System (ADS)

Covele, B.; Kotschenreuther, M.; Mahajan, S.; Valanju, P.; Leonard, A.; Watkins, J.; Makowski, M.; Fenstermacher, M.; Si, H.

2017-08-01

The X-divertor geometry on DIII-D has demonstrated reduced particle and heat fluxes to the target, facilitating detachment onset at 10-20% lower upstream density and higher H-mode pedestal pressure than a standard divertor. SOLPS modeling suggests that this effect cannot be explained by an increase in total connection length alone, but rather by the addition of connection length specifically in the power-dissipating volume near the target, via poloidal flux expansion and flaring. However, poloidal flaring must work synergistically with divertor closure to most effectively reduce the detachment density threshold. The model also points to carbon radiation as the primary driver of power dissipation in divertors on the DIII-D floor, which is consistent with experimental observations. Sustainable divertor detachment at lower density has beneficial consequences for energy confinement and current drive efficiency for core operation, while simultaneously satisfying the exhaust requirements of the plasma-facing components.

Early wound healing of laser in situ keratomileusis-like flaps after treatment with human corneal stromal stem cells.

PubMed

Morgan, Siân R; Dooley, Erin P; Kamma-Lorger, Christina; Funderburgh, James L; Funderburgh, Martha L; Meek, Keith M

2016-02-01

To use a well-established organ culture model to investigate the effects of corneal stromal stem cells on the optical and biomechanical properties of corneal wounds after laser in situ keratomileusis (LASIK)-like flap creation. School of Optometry and Vision Sciences, Cardiff University, Cardiff, Wales, United Kingdom. Experimental study. The LASIK-like flaps were produced in sheep corneas. The flap beds were treated with corneal stromal stem cells and were then replaced and allowed to heal for different periods of up to 3 weeks in organ culture. The optical transmission of the cornea, the force required to detach the flap, and the presence of myofibroblasts near the flap bed were measured. Corneal stromal stem cell-treated flap beds were statistically significantly more transparent after 3 weeks in culture than the untreated controls. At 3 weeks, the mean force necessary to detach the flap was more than twice the force required for the respective control samples. Concurrently, there were 44% activated cells immediately below the flap margin of the controls compared with 29% in the same region of the corneal stromal stem cell-treated flaps. In this system, the presence of corneal stromal stem cells at the wound margin significantly increased the adherence of LASIK-like flaps while maintaining corneal transparency. It is postulated that this is achieved by the deposition of extracellular connective tissue similar to that found in the normal cornea and by the paucity of activated keratocytes (myofibroblasts), which are known to scatter a significant amount of the incident light. No author has a financial or proprietary interest in any material or method mentioned. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Measuring Detachment of Aspergillus niger spores from Colonies with an Atomic Force Microscope.

PubMed

Li, Xian; Zhang, Tengfei Tim; Wang, Shugang

2018-06-26

Detachment of fungal spores from moldy surfaces and the subsequent aerosolization can lead to adverse health effects. Spore aerosolization occurs when the forces for aerosolization exceed the binding forces of spores with their colonies. The threshold force to detach a spore from a growing colony remains unknown. This investigation measured the detachment of spores of Aspergillus niger from a colony using an atomic force microscope (AFM). The spores were first affixed to the cantilever of the AFM with ultraviolet curing glue, and then the colony was moved downward until the spores detached. The threshold detachment forces were inferred from the deflection of the cantilever. In addition, the spores were aerosolized in a wind tunnel by a gradual increase of the blowing air speed. The forces measured by the AFM were compared with the hydrodynamic forces for aerosolization. The AFM measurements revealed that a force of 3.27 ± 0.25 nN was required to detach a single spore from the four-day-old colony, while 1.98 ± 0.13 nN was sufficient for the 10-day-old colony. Slightly smaller detachment forces were observed by the AFM than were determined by the aerosolization tests. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Staying well and engaged when demands are high: the role of psychological detachment.

PubMed

Sonnentag, Sabine; Binnewies, Carmen; Mojza, Eva J

2010-09-01

The authors of this study examined the relation between job demands and psychological detachment from work during off-job time (i.e., mentally switching off) with psychological well-being and work engagement. They hypothesized that high job demands and low levels of psychological detachment predict poor well-being and low work engagement. They proposed that psychological detachment buffers the negative impact of high job demands on well-being and work engagement. A longitudinal study (12-month time lag) with 309 human service employees showed that high job demands predicted emotional exhaustion, psychosomatic complaints, and low work engagement over time. Psychological detachment from work during off-job time predicted emotional exhaustion and buffered the relation between job demands and an increase in psychosomatic complaints and between job demands and a decrease in work engagement. The findings of this study suggest that psychological detachment from work during off-job time is an important factor that helps to protect employee well-being and work engagement. Copyright 2010 APA, all rights reserved

FAS apoptotic inhibitory molecule 2 is a stress-induced intrinsic neuroprotective factor in the retina.

PubMed

Pawar, Mercy; Busov, Boris; Chandrasekhar, Aaruran; Yao, Jingyu; Zacks, David N; Besirli, Cagri G

2017-10-01

We report the neuroprotective role of FAS apoptotic inhibitory molecule 2 (FAIM2), an inhibitor of the FAS signaling pathway, during stress-induced photoreceptor apoptosis. Retinal detachment resulted in increased FAIM2 levels in photoreceptors with higher amounts detected at the tips of outer segments. Activation of FAS death receptor via FAS-ligand led to JNK-mediated FAIM2 phosphorylation, decreased proteasome-mediated degradation and increased association with the FAS receptor. Photoreceptor apoptosis was accelerated in Faim2 knockout mice following experimental retinal detachment. We show that FAIM2 is primarily involved in reducing stress-induced photoreceptor cell death but this effect was transient. FAIM2 was found to interact with both p53 and HSP90 following the activation of the FAS death pathway and FAIM2/HSP90 interaction was dependent on the phosphorylation of FAIM2. Lack of FAIM2 led to increased expression of proadeath genes Fas and Ripk1 in the retina under physiologic conditions. These results demonstrate that FAIM2 is an intrinsic neuroprotective factor activated by stress in photoreceptors and delays FAS-mediated photoreceptor apoptosis. Modulation of this pathway to increase FAIM2 expression may be a potential therapeutic option to prevent photoreceptor death.

Abscisic Acid Accumulation in Spinach Leaf Slices in the Presence of Penetrating and Nonpenetrating Solutes 1

PubMed Central

Creelman, Robert A.; Zeevaart, Jan A. D.

1985-01-01

Abscisic acid (ABA) accumulated in detached, wilted leaves of spinach (Spinacia oleracea L. cv Savoy Hybrid 612) and reached a maximum level within 3 to 4 hours. The increase in ABA over that found in detached turgid leaves was approximately 10-fold. The effects of water stress could be mimicked by the use of thin slices of spinach leaves incubated in the presence of 0.6 molar mannitol, a compound which causes plasmolysis (loss of turgor). About equal amounts of ABA were found both in the leaf slices and in detached leaves, whereas 2 to 4 times more ABA accumulated in the medium than in the slices. When spinach leaf slices were incubated with ethylene glycol, a compound which rapidly penetrates the cell membrane causing a decrease in the osmotic potential of the tissue and only transient loss of turgor, no ABA accumulated. Ethylene glycol was not inhibitory with respect to ABA accumulation. Spinach leaf slices incubated in both ethylene glycol and mannitol had ABA levels similar to those found when slices were incubated with mannitol alone. Increases similar to those found with mannitol also occurred when Aquacide III, a highly purified form of polyethylene glycol, was used. Aquacide III causes cytorrhysis, a situation similar to that found in wilted leaves. Thus, it appears that loss of turgor is essential for ABA accumulation. When spinach leaf slices were incubated with solutes which are supposed to disturb membrane integrity (KHSO3, 2-propanol, or KCl) no increase in ABA was observed. These data indicate that, with respect to the accumulation of ABA, mannitol caused a physical stress (loss of turgor) rather than a chemical stress (membrane damage). PMID:16664022

Abscisic Acid accumulation in spinach leaf slices in the presence of penetrating and nonpenetrating solutes.

PubMed

Creelman, R A; Zeevaart, J A

1985-01-01

Abscisic acid (ABA) accumulated in detached, wilted leaves of spinach (Spinacia oleracea L. cv Savoy Hybrid 612) and reached a maximum level within 3 to 4 hours. The increase in ABA over that found in detached turgid leaves was approximately 10-fold. The effects of water stress could be mimicked by the use of thin slices of spinach leaves incubated in the presence of 0.6 molar mannitol, a compound which causes plasmolysis (loss of turgor). About equal amounts of ABA were found both in the leaf slices and in detached leaves, whereas 2 to 4 times more ABA accumulated in the medium than in the slices. When spinach leaf slices were incubated with ethylene glycol, a compound which rapidly penetrates the cell membrane causing a decrease in the osmotic potential of the tissue and only transient loss of turgor, no ABA accumulated. Ethylene glycol was not inhibitory with respect to ABA accumulation. Spinach leaf slices incubated in both ethylene glycol and mannitol had ABA levels similar to those found when slices were incubated with mannitol alone. Increases similar to those found with mannitol also occurred when Aquacide III, a highly purified form of polyethylene glycol, was used. Aquacide III causes cytorrhysis, a situation similar to that found in wilted leaves. Thus, it appears that loss of turgor is essential for ABA accumulation.When spinach leaf slices were incubated with solutes which are supposed to disturb membrane integrity (KHSO(3), 2-propanol, or KCl) no increase in ABA was observed. These data indicate that, with respect to the accumulation of ABA, mannitol caused a physical stress (loss of turgor) rather than a chemical stress (membrane damage).

Photoreceptor cell death and rescue in retinal detachment and degenerations

PubMed Central

Murakami, Yusuke; Notomi, Shoji; Hisatomi, Toshio; Nakazawa, Toru; Ishibashi, Tatsuro; Miller, Joan W.; Vavvas, Demetrios G.

2013-01-01

Photoreceptor cell death is the ultimate cause of vision loss in various retinal disorders, including retinal detachment (RD). Photoreceptor cell death has been thought to occur mainly through apoptosis, which is the most characterized form of programmed cell death. The caspase family of cysteine proteases plays a central role for inducing apoptosis, and in experimental models of RD, dying photoreceptor cells exhibit caspase activation; however, there is a paradox that caspase inhibition alone does not provide a sufficient protection against photoreceptor cell loss, suggesting that other mechanisms of cell death are involved. Recent accumulating evidence demonstrates that non-apoptotic forms of cell death, such as autophagy and necrosis, are also regulated by specific molecular machinery, such as those mediated by autophagy-related proteins and receptor-interacting protein kinases, respectively. Here we summarize the current knowledge of cell death signaling and its roles in photoreceptor cell death after RD and other retinal degenerative diseases. A body of studies indicate that not only apoptotic but also autophagic and necrotic signaling are involved in photoreceptor cell death, and that combined targeting of these pathways may be an effective neuroprotective strategy for retinal diseases associated with photoreceptor cell loss. PMID:23994436

Detachable glass microelectrodes for recording action potentials in active moving organs.

PubMed

Barbic, Mladen; Moreno, Angel; Harris, Tim D; Kay, Matthew W

2017-06-01

Here, we describe new detachable floating glass micropipette electrode devices that provide targeted action potential recordings in active moving organs without requiring constant mechanical constraint or pharmacological inhibition of tissue motion. The technology is based on the concept of a glass micropipette electrode that is held firmly during cell targeting and intracellular insertion, after which a 100-µg glass microelectrode, a "microdevice," is gently released to remain within the moving organ. The microdevices provide long-term recordings of action potentials, even during millimeter-scale movement of tissue in which the device is embedded. We demonstrate two different glass micropipette electrode holding and detachment designs appropriate for the heart (sharp glass microdevices for cardiac myocytes in rats, guinea pigs, and humans) and the brain (patch glass microdevices for neurons in rats). We explain how microdevices enable measurements of multiple cells within a moving organ that are typically difficult with other technologies. Using sharp microdevices, action potential duration was monitored continuously for 15 min in unconstrained perfused hearts during global ischemia-reperfusion, providing beat-to-beat measurements of changes in action potential duration. Action potentials from neurons in the hippocampus of anesthetized rats were measured with patch microdevices, which provided stable base potentials during long-term recordings. Our results demonstrate that detachable microdevices are an elegant and robust tool to record electrical activity with high temporal resolution and cellular level localization without disturbing the physiological working conditions of the organ. NEW & NOTEWORTHY Cellular action potential measurements within tissue using glass micropipette electrodes usually require tissue immobilization, potentially influencing the physiological relevance of the measurement. Here, we addressed this limitation with novel 100-µg detachable glass microelectrodes that can be precisely positioned to provide long-term measurements of action potential duration during unconstrained tissue movement. Copyright © 2017 the American Physiological Society.

Bilateral Posterior Scleritis Associated with Giant Cell Arteritis: A Case Report.

PubMed

Erdogan, Mehmet; Sayin, Nihat; Yıldız Ekinci, Dilbade; Bayramoglu, Sadik

2017-09-15

To report a case of bilateral posterior scleritis associated with giant cell arteritis Case Report: A 62-year-old female patient presented with bilateral progressive vision loss was diagnosed with bilateral posterior scleritis. According to clinical signs and symptoms and laboratory testing, Giant cell arteritis was also diagnosed. Within 8 weeks of the corticosteroid treatment, the serous retinal detachments completely resolved and choroidal thickness decreased in both eyes. Visual acuity increased, and the symtoms related to Giant cell arteritis improved. Posterior scleritis is an inflammatory disease that may be associated with many autoimmune systemic diseases. GCA should be thought of particularly in patients over the age of 50 with bilateral involvement, and a relevant detailed history should be obtained for early and correct diagnosis and treatment.

What controls deformation in a bent three-dimensional orogen? An example from the Bolivian Andes

NASA Astrophysics Data System (ADS)

Kaislaniemi, L.; Whipp, D. M., Jr.

2017-12-01

The width of orogens is thought to be affected by both erosional intensity and strength of the rocks. Along-strike variation of the orogen width can be expected to reflect shifts in these factors. An example of such variation can be found around the Bolivian orocline, which is a change in the orientation of the central Andes, in central Bolivia, from N-S south of 18°S to roughly NW-SE in the north. This bend coincides with 50% reduction in the width of the orogen east of the Altiplano, an approximately eight-fold increase in the annual precipitation, and the presence of a basement arch that reduces the thickness of relatively weak Paleozoic sediments upon which the orogen detaches. This has led to uncertainty about whether the growth of the orogen is controlled primarily by climate (erosion) or tectonics (strength of the basal detachment). We study deformation in a segmented orogen using 3D geodynamic models to understand how along-strike variations in rainfall and basal detachment strength affect orogen deformation and growth of the frontal part of the Andean fold-and-thrust belt (FTB). We calculate the visco-plastic deformation in the retro-wedge of an Andean-style orogen using the finite element software DOUAR (Braun et al. 2008) coupled to the surface process model FastScape (Braun & Willett 2013). The model design includes the basement, the Altiplano, and the FTB east of the plateau. A weak basal detachment zone is prescribed. Strain softening allows development of new faults and free evolution of the detachment zone. The effects of varying rock strength and varying precipitation are considered to determine the primary control(s) on the geometry and evolution of curved orogens. Results show that both increased precipitation and stronger detachment zone can explain differences in the width of the FTB, as reflected in the topography. These factors, however, lead to different structural evolution of the orogen: Weak basal detachment zone promotes growth of the FTB towards the foreland, whereas strong basal detachment keeps the deformation nearer to the plateau. Increased precipitation causes strong localization of the frontal thrust and no internal deformation in the foreland or near the plateau. Strike-slip faults are produced by variation in detachment zone strength, but not by shifts in precipitation rates.

Psychological detachment as moderator between psychosocial work conditions and low back pain development.

PubMed

Mierswa, Tobias; Kellmann, Michael

2017-03-30

Recovery processes in leisure time influence the effect of psychosocial work factors on health issues. However, this function of recovery has been neglected in research regarding the influence of work-related risk factors on low back pain (LBP) development. The aim of this prospective study was to examine the function of psychological detachment - a relevant recovery experience - concerning the influence of psychosocial work factors on LBP development. A moderating function of detachment for the interplay of work factors and LBP was assumed. Sixty pain-free administrative employees of German universities completed an online survey 3 times during a 6-month period. Generalized estimating equations were used to estimate risk-factors of LBP. Analyses revealed an increased chance of LBP development for smokers and a decreasing chance when work resources were high. Detachment had no direct influence on LBP development, although it moderated the influence of work stressors and work resources on LBP. On the one hand, high detachment values seem to protect against an increased chance of LBP development when employees were confronted with high work stressors, while on the other hand high detachment values enhance the protective effect of high work resources. The results indicated a moderating role of detachment concerning the influence of psychosocial work factors on LBP development. Therefore, it is necessary to include recovery processes in future research regarding LBP development and consequently in LBP prevention concepts. Int J Occup Med Environ Health 2017;30(2):313-327. This work is available in Open Access model and licensed under a CC BY-NC 3.0 PL license.

Microphysical Modeling of Titan's Detached Haze Layer in a 3D GCM

NASA Astrophysics Data System (ADS)

Larson, Erik J.; Toon, Owen B.; West, Robert A.; Friedson, A. James

2015-11-01

We investigate the formation and seasonal cycle of the detached haze layer in Titan’s upper atmosphere using a 3D GCM with coupled aerosol microphysics. The base of the detached haze layer is defined by a local minimum in the vertical extinction profile. The detached haze is seen at all latitudes including the south pole as seen in Cassini images from 2005-2012. The layer merges into the winter polar haze at high latitudes where the Hadley circulation carries the particles downward. The hemisphere in which the haze merges with the polar haze varies with season. We find that the base of the detached haze layer occurs where there is a near balance between vertical winds and particle fall velocities. Generally the vertical variation of particle concentration in the detached haze region is simply controlled by sedimentation, so the concentration and the extinction vary roughly in proportion to air density. This variation explains why the upper part of the main haze layer, and the bulk of the detached haze layer follow exponential profiles. However, the shape of the profile is modified in regions where the vertical wind velocity is comparable to the particle fall velocity. Our simulations closely match the period when the base of the detached layer in the tropics is observed to begin its seasonal drop in altitude, and the total range of the altitude drop. However, the simulations have the base of the detached layer about 100 km lower than observed, and the time for the base to descend is slower in the simulations than observed. These differences may point to the model having somewhat lower vertical winds than occur on Titan, or somewhat too large of particle sizes, or some combination of both. Our model is consistent with a dynamical origin for the detached haze rather than a chemical or microphysical one. This balance between the vertical wind and particle fall velocities occurs throughout the summer hemisphere and tropics. The particle concentration gradients that are established in the summer hemisphere are transported to the winter hemisphere by meridional winds from the overturning Hadley cell. Our model is consistent with the disappearance of the detached haze layer in early 2014.

A smart polymeric platform for multistage nucleus-targeted anticancer drug delivery.

PubMed

Zhong, Jiaju; Li, Lian; Zhu, Xi; Guan, Shan; Yang, Qingqing; Zhou, Zhou; Zhang, Zhirong; Huang, Yuan

2015-10-01

Tumor cell nucleus-targeted delivery of antitumor agents is of great interest in cancer therapy, since the nucleus is one of the most frequent targets of drug action. Here we report a smart polymeric conjugate platform, which utilizes stimulus-responsive strategies to achieve multistage nuclear drug delivery upon systemic administration. The conjugates composed of a backbone based on N-(2-hydroxypropyl) methacrylamide (HPMA) copolymer and detachable nucleus transport sub-units that sensitive to lysosomal enzyme. The sub-units possess a biforked structure with one end conjugated with the model drug, H1 peptide, and the other end conjugated with a novel pH-responsive targeting peptide (R8NLS) that combining the strength of cell penetrating peptide and nuclear localization sequence. The conjugates exhibited prolonged circulation time and excellent tumor homing ability. And the activation of R8NLS in acidic tumor microenvironment facilitated tissue penetration and cellular internalization. Once internalized into the cell, the sub-units were unleashed for nuclear transport through nuclear pore complex. The unique features resulted in 50-fold increase of nuclear drug accumulation relative to the original polymer-drug conjugates in vitro, and excellent in vivo nuclear drug delivery efficiency. Our report provides a strategy in systemic nuclear drug delivery by combining the microenvironment-responsive structure and detachable sub-units. Copyright © 2015 Elsevier Ltd. All rights reserved.

Late Cenozoic cooling history of the central Menderes Massif and the contribution of erosion to rock exhumation during active continental extension

NASA Astrophysics Data System (ADS)

Nilius, Nils-Peter; Wölfler, Andreas; Heineke, Caroline; Glotzbach, Christoph; Hetzel, Ralf; Hampel, Andrea; Akal, Cüneyt; Dunkl, István

2017-04-01

The Menderes Massif constitutes the western part of the Anatolide belt in western Turkey and experienced a prolonged history of post-orogenic extension. A large amount of the extension was accommodated by the two oppositely dipping Gediz and Büyük Menderes detachment faults, which led to the exhumation of the central Menderes Massif (Gessner et al., 2013). Previous studies proposed a synchronous, bivergent exhumation of the central Menderes Massif since the Miocene (Gessner et al., 2001), although only the evolution of the north-dipping Gediz detachment is well constrained (Buscher et al., 2013). Detailed structural and thermochronological investigations from the south-dipping Büyük Menderes detachment have still been missing. Here we present results from different thermochronometers, which constrain the cooling and exhumation history of footwall and hanging wall rocks of the Büyük Menderes detachment. Our new zircon and apatite (U-Th)/He and fission track ages of footwall rocks from the Büyük Menderes detachment document two phases of increased cooling and exhumation (Wölfler et al., in revision). The first episode of increased footwall exhumation ( 0.9 km/Myr) occurred during the middle Miocene, followed by a second phase during latest Miocene and Pliocene ( 1.0 km/Myr). Apatite fission track ages yield a slip rate for the Pliocene movement along the Büyük Menderes detachment of 3.0 (+1.1/-0.6) km/Myr. Thermochronological data of hanging wall units reflect a slow phase of exhumation ( 0.2 km/Myr) in the late Oligocene and an increased exhumation rate of 1.0 km/Myr during the early to middle Miocene, when hanging wall units cooled below 80 °C. In comparison with the Gediz detachment, our thermochronological data from the Büyük Menderes detachment confirms the concurrent activity of both detachments during the late Miocene and Pliocene. With respect to the relative importance of normal faulting and erosion to rock exhumation, a comparison with 10Be erosion rates from catchments in the exposed footwall of the Büyük Menderes detachment indicates that erosion has contributed 10-40% to the exhumation of metamorphic rocks beneath the detachment. Our finding underlines that the contribution of erosion to rock exhumation cannot be neglected in regions of active continental extension. References Buscher, J.T., Hampel, A., Hetzel, R., Dunkl, I, Glotzbach, C., Struffert, A., Akal, C., Rätz, M. 2013. Quantifying rates of detachment faulting and erosion in the central Menderes Massif (western Turkey) by thermochronology and cosmogenic 10Be. J. Geol. Soc. London. 170, 669-683. Gessner, K., Ring, U., Johnson, C., Hetzel, R., Passchier, C.W., Güngör, T., 2001. An active bivergent rolling-hinge detachment system: Central Menderes metamorphic core complex in western Turkey. Geology 29, 611-614. Gessner, K., Gallardo, L.A., Markwitz, V., Ring, U., Thomson, S.N., 2013. What caused the denudation of the Menderes Massif: Review of crustal evolution, lithosphere structure, and dynamic topography in southwest Turkey. Gondwana Research 24, 243-274. Wölfler, A., Glotzbach, C., Heineke, C., Nilius, N.P., Hetzel, R., Hampel, A., Akal, C., Dunkl, I., Christl, M. (manuscript in revision for Tectonophysics). Late Cenozoic cooling history of the central Menderes Massif: timing and slip rate of the Büyük Menderes detachment and the relative contribution of normal faulting and erosion to rock exhumation.

Stochastic mechanics of loose boundary particle transport in turbulent flow

NASA Astrophysics Data System (ADS)

Dey, Subhasish; Ali, Sk Zeeshan

2017-05-01

In a turbulent wall shear flow, we explore, for the first time, the stochastic mechanics of loose boundary particle transport, having variable particle protrusions due to various cohesionless particle packing densities. The mean transport probabilities in contact and detachment modes are obtained. The mean transport probabilities in these modes as a function of Shields number (nondimensional fluid induced shear stress at the boundary) for different relative particle sizes (ratio of boundary roughness height to target particle diameter) and shear Reynolds numbers (ratio of fluid inertia to viscous damping) are presented. The transport probability in contact mode increases with an increase in Shields number attaining a peak and then decreases, while that in detachment mode increases monotonically. For the hydraulically transitional and rough flow regimes, the transport probability curves in contact mode for a given relative particle size of greater than or equal to unity attain their peaks corresponding to the averaged critical Shields numbers, from where the transport probability curves in detachment mode initiate. At an inception of particle transport, the mean probabilities in both the modes increase feebly with an increase in shear Reynolds number. Further, for a given particle size, the mean probability in contact mode increases with a decrease in critical Shields number attaining a critical value and then increases. However, the mean probability in detachment mode increases with a decrease in critical Shields number.

Tetraspanin CD151 regulates alpha6beta1 integrin adhesion strengthening

NASA Technical Reports Server (NTRS)

Lammerding, Jan; Kazarov, Alexander R.; Huang, Hayden; Lee, Richard T.; Hemler, Martin E.

2003-01-01

The tetraspanin CD151 molecule associates specifically with laminin-binding integrins, including alpha6beta1. To probe strength of alpha6beta1-dependent adhesion to laminin-1, defined forces (0-1.5 nN) were applied to magnetic laminin-coated microbeads bound to NIH 3T3 cells. For NIH 3T3 cells bearing wild-type CD151, adhesion strengthening was observed, as bead detachment became more difficult over time. In contrast, mutant CD151 (with the C-terminal region replaced) showed impaired adhesion strengthening. Static cell adhesion to laminin-1, and detachment of beads coated with fibronectin or anti-alpha6 antibody were all unaffected by CD151 mutation. Hence, CD151 plays a key role in selectively strengthening alpha6beta1 integrin-mediated adhesion to laminin-1.

Keratocytes Generate Traction Forces in Two PhasesV⃞

PubMed Central

Burton, Kevin; Park, Jung H.; Taylor, D. Lansing

1999-01-01

Forces generated by goldfish keratocytes and Swiss 3T3 fibroblasts have been measured with nanonewton precision and submicrometer spatial resolution. Differential interference contrast microscopy was used to visualize deformations produced by traction forces in elastic substrata, and interference reflection microscopy revealed sites of cell-substratum adhesions. Force ranged from a few nanonewtons at submicrometer spots under the lamellipodium to several hundred nanonewtons under the cell body. As cells moved forward, centripetal forces were applied by lamellipodia at sites that remained stationary on the substratum. Force increased and abruptly became lateral at the boundary of the lamellipodium and the cell body. When the cell retracted at its posterior margin, cell-substratum contact area decreased more rapidly than force, so that stress (force divided by area) increased as the cell pulled away. An increase in lateral force was associated with widening of the cell body. These mechanical data suggest an integrated, two-phase mechanism of cell motility: (1) low forces in the lamellipodium are applied in the direction of cortical flow and cause the cell body to be pulled forward; and (2) a component of force at the flanks pulls the rear margins forward toward the advancing cell body, whereas a large lateral component contributes to detachment of adhesions without greatly perturbing forward movement. PMID:10564269

Keratocytes generate traction forces in two phases.

PubMed

Burton, K; Park, J H; Taylor, D L

1999-11-01

Forces generated by goldfish keratocytes and Swiss 3T3 fibroblasts have been measured with nanonewton precision and submicrometer spatial resolution. Differential interference contrast microscopy was used to visualize deformations produced by traction forces in elastic substrata, and interference reflection microscopy revealed sites of cell-substratum adhesions. Force ranged from a few nanonewtons at submicrometer spots under the lamellipodium to several hundred nanonewtons under the cell body. As cells moved forward, centripetal forces were applied by lamellipodia at sites that remained stationary on the substratum. Force increased and abruptly became lateral at the boundary of the lamellipodium and the cell body. When the cell retracted at its posterior margin, cell-substratum contact area decreased more rapidly than force, so that stress (force divided by area) increased as the cell pulled away. An increase in lateral force was associated with widening of the cell body. These mechanical data suggest an integrated, two-phase mechanism of cell motility: (1) low forces in the lamellipodium are applied in the direction of cortical flow and cause the cell body to be pulled forward; and (2) a component of force at the flanks pulls the rear margins forward toward the advancing cell body, whereas a large lateral component contributes to detachment of adhesions without greatly perturbing forward movement.

Differential accumulation of dehydrins in response to water stress for hybrid and common bermudagrass genotypes differing in drought tolerance.

PubMed

Hu, Longxing; Wang, Zhaolong; Du, Hongmei; Huang, Bingru

2010-01-15

Expression of dehydrin proteins may be induced or enhanced by environmental stresses that lead to cell dehydration. The objective of the this study was to investigate genetic variation in dehydrin protein accumulation in response to drought stress of whole-plants or dehydration of detached leaves and to identify dehydrins differentially expressed in bermudagrass (Cynodon spp.) genotypes differing in drought tolerance. Plants of four hybrid bermudagrass (Cynodondactylon L. xCynodontransvaalensis L.) ('Tifway', 'Tifdwarf', 'Tifeagle', 'Kan1') and four common bermudagrass (Cynodon dactylon) ('C299', 'Sportbermuda', 'H10', and 'H19') genotypes were subjected to 14d of drought stress and detached leaves of two genotypes were exposed to dehydration in growth chambers. Turf quality and leaf relative water content (RWC) decreased while electrolyte leakage (EL) increased during whole-plant drought stress for all genotypes, with more pronounced changes in each parameter for 'C299' and 'Tifeagle' than those for other genotypes ('Tifway', 'Kan 1', 'Sportbermuda', 'H10', and H19'), suggesting that the former two genotypes were more sensitive to drought stress than the other genotypes. During dehydration of detached leaves, relative water loss rate (RWL) was significantly lower in drought-tolerant 'Tifway' than in drought-sensitive 'C299'. Immunoblotting analysis indicated that no dehydrin polypeptides were detected in all genotypes under well-watered conditions. A 24-kDa polypeptide was detected in 'C299' at 6 d of drought, but not in the other genotypes. The dehydrin polypeptides of about 14-74kDa accumulated at 10d of drought stress and in a range of RWL for detached leaves, and two dehydrins (31 and 40kDa) exhibited differential accumulation in the drought-sensitive 'C299' and tolerant 'Tifway', as demonstrated by the whole-plant drought responses. The 31-kDa dehydrin polypeptide was present only in 'Tifway' and 'H19' at 10d of drought stress, and accumulated with the increasing RWL in detached leaves of 'Tifway'. The expression level of 40-kDa dehydrin polypeptides was greater in 'Tifway'' than in 'C299' at the same level of water deficit (from 10% to 65% RWL). These results indicated that the accumulation of 31- and 40-kDa dehydrins may contribute to drought or dehydration tolerance in warm-season bermudagrass.

Turbulent Simulations of Divertor Detachment Based On BOUT + + Framework

NASA Astrophysics Data System (ADS)

Chen, Bin; Xu, Xueqiao; Xia, Tianyang; Ye, Minyou

2015-11-01

China Fusion Engineering Testing Reactor is under conceptual design, acting as a bridge between ITER and DEMO. The detached divertor operation offers great promise for a reduction of heat flux onto divertor target plates for acceptable erosion. Therefore, a density scan is performed via an increase of D2 gas puffing rates in the range of 0 . 0 ~ 5 . 0 ×1023s-1 by using the B2-Eirene/SOLPS 5.0 code package to study the heat flux control and impurity screening property. As the density increases, it shows a gradually change of the divertor operation status, from low-recycling regime to high-recycling regime and finally to detachment. Significant radiation loss inside the confined plasma in the divertor region during detachment leads to strong parallel density and temperature gradients. Based on the SOLPS simulations, BOUT + + simulations will be presented to investigate the stability and turbulent transport under divertor plasma detachment, particularly the strong parallel gradient driven instabilities and enhanced plasma turbulence to spread heat flux over larger surface areas. The correlation between outer mid-plane and divertor turbulence and the related transport will be analyzed. Prepared by LLNL under Contract DE-AC52-07NA27344. LLNL-ABS-675075.

The Role of an Electric Field in the Formation of a Detached Regime in Tokamak Plasma

NASA Astrophysics Data System (ADS)

Senichenkov, I.; Kaveeva, E.; Rozhansky, V.; Sytova, E.; Veselova, I.; Voskoboynikov, S.; Coster, D.

2018-03-01

Modeling of the transition to the detachment of ASDEX Upgrade tokamak plasma with increasing density is performed using the SOLPS-ITER numerical code with a self-consistent account of drifts and currents. Their role in plasma redistribution both in the confinement region and in the scrape-off layer (SOL) is investigated. The mechanism of high field side high-density formation in the SOL in the course of detachment is suggested. In the full detachment regime, when the cold plasma region expands above the X-point and reaches closed magnetic-flux surfaces, plasma perturbation in a confined region may lead to a change in the confinement regime.

Glaucoma with Descemet's membrane detachment in five horses.

PubMed

Henriksen, Michala de Linde; La Croix, Noelle; Wilkie, David A; Lassaline-Utter, Mary; Brantman, Karen R; Beamer, Gillian L; Teixeira, Leandro B C; Dubielzig, Richard R

2017-05-01

To describe the clinical and histopathologic features of glaucoma associated with Descemet's membrane (DM) detachment in five horses without prior history of intraocular surgery. Three Appaloosa horses and two Thoroughbreds were included in this study. The affected horses ranged in age from 16 to 27 years and presented with severe diffuse corneal edema. Five eyes were enucleated due to intraocular hypertension and/or chronic corneal ulceration. The enucleated globes were evaluated by the Comparative Ocular Pathology Laboratory of Wisconsin (COPLOW). Each globe was routinely processed for histopathology and analyzed by light microscopy. A histologic diagnosis of glaucoma was reached by demonstrating a loss of optic nerve axonal tissue by measuring neurofilament-immunopositive axons with automated image analysis software. All five horses presented with unilateral severe diffuse corneal edema that had developed between 2 and 16 weeks prior to enucleation. Intraocular pressures for the affected eyes were between 9 and 87 mmHg prior to enucleation. Descemet's membrane detachment was identified histopathologically in all five globes (5/5, 100%). All five eyes had an avascular spindle cell proliferation filling the space between the displaced peripheral DM and the corneal stroma. Neurofilament immunostaining revealed axonal loss consistent with glaucoma. Equine glaucoma may be associated with Descemet's membrane detachment. This detachment and glaucoma is a possible differential diagnosis for severe equine corneal edema. In this case series, an eye with a DM detachment had a poor prognosis for retention. © 2016 American College of Veterinary Ophthalmologists.

GENE EXPRESSION PROFILING OF HUMAN LIVER CARCINOMA (HepG2) CELLS EXPOSED TO THE MARINE TOXIN OKADAIC ACID

PubMed Central

Fieber, Lynne A.; Greer, Justin B.; Guo, Fujiang; Crawford, Douglas C.; Rein, Kathleen S.

2012-01-01

The marine toxin, okadaic acid (OA) is produced by dinoflagellates of the genera Prorocentrum and Dinophysis and is the causative agent of the syndrome known as diarrheic shellfish poisoning (DSP). In addition, OA acts as both a tumor promoter, attributed to OA-induced inhibition of protein phosphatases as well as an inducer of apoptosis. To better understand the potentially divergent toxicological profile of OA, the concentration dependent cytotoxicity and alterations in gene expression on the human liver tumor cell line HepG2 upon OA exposure were determined using RNA microarrays, DNA fragmentation, and cell proliferation assays as well as determinations of cell detachment and cell death in different concentrations of OA. mRNA expression was quantified for approximately 15,000 genes. Cell attachment and proliferation were both negatively correlated with OA concentration. Detached cells displayed necrotic DNA signatures but apoptosis also was broadly observed. Data suggest that OA has a concentration dependent effect on cell cycle, which might explain the divergent effects that at low concentration OA stimulates genes involved in the cell cycle and at high concentrations it stimulates apoptosis. PMID:23172983

Long-term effects of short-term retinal bleb detachments in rabbits.

PubMed

Ivert, Lena; Kjeldbye, Hild; Gouras, Peter

2002-03-01

To examine the effects of saline-induced bleb detachments in rabbit retina. Retinal bleb detachments were produced by the injection of 50 microl of balanced salt solution (BSS) into the subretinal space of one eye of each of six rabbits using a glass pipette with a flat tip, 50 microm in diameter. The retina was examined by biomicroscopy, scanning laser ophthalmoscopy (SLO), auto-fluorescence and simultaneous fluorescein and indocyanine green (ICG) angiography. Histological examination was carried out at 1, 2, 3 and 4 months after surgery. All rabbits showed leakage of fluorescein for at least a day after detachment, but within 1 month the leakage ceased. ICG staining developed gradually at the level of the RPE or Bruch's membrane near sites of previous staining. Lipofuscin fluorescence also developed gradually around areas of staining. Histology revealed the source of the excessive lipofuscin to be in the RPE layer, especially in cells migrating away from Bruch's membrane. Short-term bleb detachments cause a transient breakdown in the blood-retinal barrier, long-term ICG staining at or deep to the RPE layer, hyperlipofuscinosis and migration of the RPE. The abnormal lipofuscin accumulation is apparent on fluorescence ophthalmoscopy and can be confused with markers such as green fluorescent protein.

Prevalence of early and late stages of physiologic PVD in emmetropic elderly population.

PubMed

Schwab, Christoph; Ivastinovic, Domagoj; Borkenstein, Andreas; Lackner, Eva-Maria; Wedrich, Andreas; Velikay-Parel, Michaela

2012-05-01

To investigate the early and late stages of posterior vitreous detachment (PVD) in the foveal area in correlation with age and gender. Three hundred and thirty-five emmetropic eyes of 271 Caucasian patients (216 women/119 men) were examined by optical coherence tomography (OCT) and ultrasound (US). Eyes were classified into groups according to the patients age (up to 69.9; 70-74.9; 75-79.9; over 80 years) and to the clinical findings [Vitreous state: Detached in US; Detached in OCT; Foveal adhesion (FA); Attached vitreous]. The mean age was 76 ± 8 ranging from 44 to 89 years in female and 72 ± 10 ranging from 46 to 87 years in male subjects. The vitreous was attached in 32% of all eyes, 18.5% had FA, 18.5% were detached in OCT and 68% were detached in US. While prevalence of FA decreases with increasing age, OCT-diagnosed detachments did not change significantly with age. Between the ages of 70 and 75, an increase in PVD rates occurred. The prevalence of PVD was similar in both genders. Women were significantly older than men in the late-stage PVD in the eyes. The use of OCT and US enabled us to detect a partial or total PVD in 80% of the eyes. A sudden increase in late-stage PVD between the ages of 70 and 75 was observed, correlating with the reported age prevalence of various macular diseases. In contrast to myopics, both genders of elderly emmetropics have a similar prevalence of PVD. © 2011 The Authors. Acta Ophthalmologica © 2011 Acta Ophthalmologica Scandinavica Foundation.

Treponema denticola Outer Membrane Enhances the Phagocytosis of Collagen-Coated Beads by Gingival Fibroblasts

PubMed Central

Battikhi, Tulin; Lee, Wilson; McCulloch, Christopher A. G.; Ellen, Richard P.

1999-01-01

Human gingival fibroblasts (HGFs) degrade collagen fibrils in physiological processes by phagocytosis. Since Treponema denticola outer membrane (OM) extract perturbs actin filaments, important structures in phagocytosis, we determined whether the OM affects collagen phagocytosis in vitro by HGFs. Phagocytosis was measured by flow cytometric assessment of internalized collagen-coated fluorescent latex beads. Confluent HGFs pretreated with T. denticola ATCC 35405 OM exhibited an increase in the percentage of collagen phagocytic cells (phagocytosis index [PI]) and in the number of beads per phagocytosing cell (phagocytic capacity [PC]) compared with untreated controls. The enhancement was swift (within 15 min) and was still evident after 1 day. PI and PC of HGFs for bovine serum albumin (BSA)-coated beads were also increased, indicating a global increase in phagocytic processes. These results contrasted those for control OM from Veillonella atypica ATCC 17744, which decreased phagocytosis. The T. denticola OM-induced increase in bead uptake was eliminated by heating the OM and by depolymerization of actin filaments by cytochalasin D treatment of HGFs. Fluid-phase accumulation of lucifer yellow was enhanced in a saturable, concentration-dependent, transient manner by the T. denticola OM. Our findings were not due to HGF detachment or cytotoxicity in response to the T. denticola OM treatment since the HGFs exhibited minimal detachment from the substratum; they did not take up propidium iodide; and there was no change in their size, granularity, or content of sub-G1 DNA. We conclude that a heat-sensitive component(s) in T. denticola OM extract stimulates collagen phagocytosis and other endocytic processes such as nonspecific phagocytosis and pinocytosis by HGFs. PMID:10024564

Effects of Near Soil Surface Characteristics on the Soil Detachment Process in a Chronological Series of Vegetation Restoration

NASA Astrophysics Data System (ADS)

Wang, Bing

2017-04-01

The effects of near soil surface characteristics on the soil detachment process might be different at different stages of vegetation restoration. This study was performed to investigate the effects of the near soil surface factors of plant litter, biological soil crusts (BSCs), dead roots and live roots on the soil detachment process by overland flow at different stages of restoration. Soil samples (1 m long, 0.1 m wide, and 0.05 m high) under four treatment conditions were collected from 1-yr-old and 24-yr-old natural grasslands and subjected to flow scouring under five different shear stresses ranging from 5.3 to 14.6 Pa. The results indicated that the effects of near soil surface characteristics on soil detachment were substantial during the process of vegetation restoration. The total reduction in the soil detachment capacity of the 1-yr-old grassland was 98.1%, and of this total, 7.9%, 30.0% and 60.2% was attributed to the litter, BSCs and plant roots, respectively. In the 24-yr-old grassland, the soil detachment capacity decreased by 99.0%, of which 13.2%, 23.5% and 62.3% was caused by the litter, BSCs and plant roots, respectively. Combined with the previously published data of a 7-yr-old grassland, the influence of plant litter on soil detachment was demonstrated to increase with restoration time, but soil detachment was also affected by the litter type and composition. The role of BSCs was greater than that of plant litter in reducing soil detachment during the early stages of vegetation recovery. However, its contribution weakened with time since restoration. The influence of plant roots accounted for at least half or up to two-thirds of the total near soil surface factors, of which more than 72.6% was attributed to the physical binding effects of the roots. The chemical bonding effect of the roots increased with time since restoration and was greater than the effect of the litter on soil detachment in the late stages of vegetation restoration. The correction coefficients of near soil surface characteristics for rill erodibility were provided for the Water Erosion Prediction Project (WEPP) model.

Gaining insights into interrill soil erosion processes using rare earth element tracers

USDA-ARS?s Scientific Manuscript database

Increasing interest in developing process-based erosion models requires better understanding of the relationships among soil detachment, transportation, and deposition. The objectives are to 1) identify the limiting process between soil detachment and sediment transport for interrill erosion, 2) und...

Increased heat dissipation with the X-divertor geometry facilitating detachment onset at lower density in DIII-D

DOE PAGES

Covele, Brent; Kotschenreuther, M.; Mahajan, S.; ...

2017-06-23

The X-Divertor geometry on DIII-D has demonstrated reduced particle and heat fluxes to the target, facilitating detachment onset at ~20% lower upstream density and higher H-mode pedestal pressure than a standard divertor. SOLPS modeling suggests that this effect cannot be explained by an increase in total connection length alone, but rather by the addition of connection length specifically in the power-dissipating volume near the target, via poloidal flux expansion and flaring. But, poloidal flaring must work synergistically with divertor closure to most effectively reduce the detachment density threshold. Furthermore, the model also points to carbon radiation as the primary drivermore » of power dissipation in divertors on the DIII-D floor, which is consistent with experimental observations. Sustainable divertor detachment at lower density has beneficial consequences for energy confinement and current drive efficiency in the core for advanced tokamak (AT) operation, while simultaneously satisfying the exhaust requirements of the plasma-facing components.« less

[Photosynthetic characteristics of Cuscuta japonica and its hosts during parasitization and after detachment].

PubMed

Wang, Dong; Hu, Fei; Chen, Yu-Fen; Yang, Jun; Kong, Chui-Hua

2007-08-01

The study on the photosynthetic characteristics of Cuscuta japonica and its hosts showed that there was a negative correlation between the photosynthetic pigment content (PPC) of C. japonica and its hosts. The PPC increased in the C. japonica-preferred hosts' parasitized and neighboring leaves, but decreased in its less preferred hosts' parasitized and neighboring leaves. The leaves parasitized by C. japonica and their neighboring far from the parasitized ones had a lowered net photosynthesis rate P(n), and the decreasing order accorded with that of parasitization. The decrease of P(n) for C. japonica-less preferred hosts was mainly due to the stomatal factors, but that for the preferred hosts was regulated by multi-factors. Under light, the PPC of C. japonica detached from preferred hosts increased faster than that of C. japonica detached from less preferred hosts, but the dry matter decrease was in adverse. In dark, however, the changes in PPC and dry matter content of C. japonica were not significant, whatever hosts it was detached from.

Increased heat dissipation with the X-divertor geometry facilitating detachment onset at lower density in DIII-D

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

Covele, Brent; Kotschenreuther, M.; Mahajan, S.

The X-Divertor geometry on DIII-D has demonstrated reduced particle and heat fluxes to the target, facilitating detachment onset at ~20% lower upstream density and higher H-mode pedestal pressure than a standard divertor. SOLPS modeling suggests that this effect cannot be explained by an increase in total connection length alone, but rather by the addition of connection length specifically in the power-dissipating volume near the target, via poloidal flux expansion and flaring. But, poloidal flaring must work synergistically with divertor closure to most effectively reduce the detachment density threshold. Furthermore, the model also points to carbon radiation as the primary drivermore » of power dissipation in divertors on the DIII-D floor, which is consistent with experimental observations. Sustainable divertor detachment at lower density has beneficial consequences for energy confinement and current drive efficiency in the core for advanced tokamak (AT) operation, while simultaneously satisfying the exhaust requirements of the plasma-facing components.« less

Effect of 3D magnetic perturbations on divertor conditions and detachment in tokamak and stellarator

DOE PAGES

Ahn, J. -W.; Briesemester, A. R.; Kobayashi, M.; ...

2017-06-22

Enhanced perpendicular heat and momentum transport induces parallel pressure loss leading to divertor detachment, which can be produced by the increase of density in 2D tokamaks. However, in the 3D configurations such as tokamaks with 3D fields and stellarators, the fraction of perpendicular transport can be higher even in a lower density regime, which could lead to the early transition to detachment without passing through the high-recycling regime. 3D fields applied to the limiter tokamak plasmas produce edge stochastic layers close to the last closed flux surface (LCFS), which can allow for enhanced perpendicular transport and indeed the absence ofmore » high recycling regime and early detachment have been observed in TEXTOR and Tore Supra. However, in the X-point divertor tokamaks with the applied 3D fields, the parallel transport is still dominant and the detachment facilitation has not been observed yet. Rather, 3D fields affected detachment adversely under certain conditions, either by preventing detachment onset as seen in DIII-D or by re-attaching the existing detached plasma as shown in NSTX. The possible way for strong 3D effects to induce access to the early detachment in divertor tokamaks appears to be via significant perpendicular loss of parallel momentum by frictional force for the counter-streaming flows between neighboring flow channels in the divertor. In principle, the adjacent lobes in the 3D divertor tokamak may generate the counter-streaming flow channels. However, an EMC3-EIRENE simulation for ITER H-mode plasmas demonstrated that screened RMP leads to significantly reduced counter-flows near the divertor target, therefore the momentum loss effect leading to detachment facilitation is expected to be small. This is consistent with the observation in LHD, which showed screening (amplification) of RMP fields in the attachment (stable detachment) case. In conclusion, work for optimal parameter window for best divertor operation scenario is needed particularly for the 3D divertor tokamak configuration.« less

Effect of 3D magnetic perturbations on divertor conditions and detachment in tokamak and stellarator

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

Ahn, J. -W.; Briesemester, A. R.; Kobayashi, M.

Enhanced perpendicular heat and momentum transport induces parallel pressure loss leading to divertor detachment, which can be produced by the increase of density in 2D tokamaks. However, in the 3D configurations such as tokamaks with 3D fields and stellarators, the fraction of perpendicular transport can be higher even in a lower density regime, which could lead to the early transition to detachment without passing through the high-recycling regime. 3D fields applied to the limiter tokamak plasmas produce edge stochastic layers close to the last closed flux surface (LCFS), which can allow for enhanced perpendicular transport and indeed the absence ofmore » high recycling regime and early detachment have been observed in TEXTOR and Tore Supra. However, in the X-point divertor tokamaks with the applied 3D fields, the parallel transport is still dominant and the detachment facilitation has not been observed yet. Rather, 3D fields affected detachment adversely under certain conditions, either by preventing detachment onset as seen in DIII-D or by re-attaching the existing detached plasma as shown in NSTX. The possible way for strong 3D effects to induce access to the early detachment in divertor tokamaks appears to be via significant perpendicular loss of parallel momentum by frictional force for the counter-streaming flows between neighboring flow channels in the divertor. In principle, the adjacent lobes in the 3D divertor tokamak may generate the counter-streaming flow channels. However, an EMC3-EIRENE simulation for ITER H-mode plasmas demonstrated that screened RMP leads to significantly reduced counter-flows near the divertor target, therefore the momentum loss effect leading to detachment facilitation is expected to be small. This is consistent with the observation in LHD, which showed screening (amplification) of RMP fields in the attachment (stable detachment) case. In conclusion, work for optimal parameter window for best divertor operation scenario is needed particularly for the 3D divertor tokamak configuration.« less

Improving the performances of H-Darrieus cross-flow turbines through proper detached end plate designs

NASA Astrophysics Data System (ADS)

Villeneuve, Thierry; Boudreau, Matthieu; Dumas, Guy; CFD Laboratory LMFN Team

2017-11-01

Previous studies on H-Darrieus cross-flow turbines have highlighted the fact that their performances are highly sensitive to the detrimental effects associated with the blades tips. Wingtip devices could be designed in order to attenuate these effects, but the benefits of such devices are always impaired by their added viscous drag since they are moving with the turbine's blades. In this context, the development of fixed and detached end plates, i.e., which are not in contact with the turbine's blades, could reduce the tip losses without the undesirable added drag of typical wingtip devices moving with the blades. The case of a single stationary blade with detached end plates has first been investigated with RANS simulations in order to understand the mechanisms responsible for the increase of the blade's lift. An analysis of the vorticity lines' dynamics provides crucial insights into the effects of the gap width between the blade and the detached end plate on the blade's loading and on the intensity of the tip vortices. Based on these observations, various configurations of detached end plates are tested on cross-flow turbines via RANS and DDES simulations. Preliminary results show that appropriate detached end plates can significantly increase the turbines' efficiency. The authors gratefully acknowledge the Natural Sciences and Engineering Research Council of Canada (NSERC) for their financial support as well as Compute Canada and Calcul Québec for their supercomputer allocations.

Numerical experiment to estimate the validity of negative ion diagnostic using photo-detachment combined with Langmuir probing

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

Oudini, N.; Sirse, N.; Ellingboe, A. R.

2015-07-15

This paper presents a critical assessment of the theory of photo-detachment diagnostic method used to probe the negative ion density and electronegativity α = n{sub -}/n{sub e}. In this method, a laser pulse is used to photo-detach all negative ions located within the electropositive channel (laser spot region). The negative ion density is estimated based on the assumption that the increase of the current collected by an electrostatic probe biased positively to the plasma is a result of only the creation of photo-detached electrons. In parallel, the background electron density and temperature are considered as constants during this diagnostics. While the numericalmore » experiments performed here show that the background electron density and temperature increase due to the formation of an electrostatic potential barrier around the electropositive channel. The time scale of potential barrier rise is about 2 ns, which is comparable to the time required to completely photo-detach the negative ions in the electropositive channel (∼3 ns). We find that neglecting the effect of the potential barrier on the background plasma leads to an erroneous determination of the negative ion density. Moreover, the background electron velocity distribution function within the electropositive channel is not Maxwellian. This is due to the acceleration of these electrons through the electrostatic potential barrier. In this work, the validity of the photo-detachment diagnostic assumptions is questioned and our results illustrate the weakness of these assumptions.« less

Feasibility studies of oncornavirus production in microcarrier cultures.

PubMed

Manousos, M; Ahmed, M; Torchio, C; Wolff, J; Shibley, G; Stephens, R; Mayyasi, S

1980-06-01

Studies conducted with virus-infected monolayer cell cultures have demonstrated the feasibility of producing several tumor-associated viruses in microcarrier (mc) cultures (Sephadex G50 beads treated with DEAE-chloride). The efficiency of cell adherence to mc varied with the cell type, the pH of the growth medium, and the stirring force applied to keep the mc in suspension. Most cells attached firmly to the mc and could not be removed easily with routine trypsinization procedures. Techniques using Enzar-T and Pronase were effective in detaching cells from mc in 10 to 15 min while retaining 95% cell viability. After detachment, Ficoll gradients were used for rapid and complete separation of viable cell suspensions from the mc. Retrovirus production in large volumes of mc cultures was investigated with periodic harvesting of growth fluids. Physical, biochemical, and biological properties of the Mason-Pfizer monkey virus and the RD114 virus recovered from the mc cultures were identical to those produced in conventional cultures. The utilization of mc has several applications in research and short-term cultures, but the as-yet-unsolved technical problems met were found to be serious limitations when attempting mass cell culturing on a long-term basis.

An investigation of cell growth and detachment from thermoresponsive physically crosslinked networks.

PubMed

Healy, Deirdre; Nash, Maria; Gorleov, Alexander; Thompson, Kerry; Dockery, Peter; Rochev, Yury

2017-11-01

The primary aim of this investigation was to determine the biocompatibility and cell culture potential of a newly designed class of thermoresponsive polymers. The attractiveness of these polymers lies in the fact that they swell rather than dissolve when the temperature is reduced below their respective lower critical solution temperature, due to the incorporation of octadecyl methacrylate (ODMA). The ODMA monomer acts as a physical crosslinker, preventing polymer dissolution upon temperature reduction. Two polymers were studied in this investigation poly(N isorpoylacrylamide (NIPAm)(99.25%)-co-ODMA(0.75%)) and poly(NIPAm(65%)-co-N-tert-butylacrylamide (NtBAm)(34.25%)-co-ODMA(0.75%)). Thin thermoresponsive films of the polymers were prepared via spin coating. 3T3 cells were then seeded on the prepared films and cell viability was assessed quantitatively through cell viability and activity assays and qualitatively by light microscopy. Cells were successfully seeded and grown on the poly(NIPAm-co-ODMA) and poly(NIPAm-co-NtBAm-co-ODMA) copolymer films after film modification with cell adhesion promoters (CAPs). Cell sheets successfully detached from the CAP coated poly(NIPAm-co-ODMA) platforms upon temperature reduction. Copyright © 2017 Elsevier B.V. All rights reserved.

Microvascular hemodynamics and in vivo evidence for the role of intercellular adhesion molecule-1 in the sequestration of infected red blood cells in a mouse model of lethal malaria.

PubMed

Kaul, D K; Liu, X D; Nagel, R L; Shear, H L

1998-02-01

The cytoadherence of infected red blood cells (IRBCs) to the vascular endothelium is the major cause of IRBC sequestration and vessel blockage in the cerebral form of human malaria. Among the rodent models of malaria, Plasmodium yoelii 17XL-infected mice show many similarities with the human cerebral malaria caused by P. falciparum. In both, the sequestration of IRBCs in the brain vessels is secondary to the cytoadherence of IRBCs to the vascular endothelium. Similar to P. falciparum infection in the human but in contrast to P. berghei ANKA infection in mice, P. yoelii 17XL results in little, if any, accumulation of monocytes in the brain. In vivo microcirculatory studies reported here were designed to further understand the hemodynamic aspects and mechanisms underlying cytoadherence of IRBCs in the P. yoelii model using the easily accessible cremaster muscle vasculature. The results show significant decreases in arteriovenous red blood cell velocities (Vrbc) and wall shear rates in the microcirculation of P. yoelii-infected mice, with a maximal decrease occurring in small-diameter postcapillary venules, the main sites of cytoadherence. This reflects contributions from IRBC cytoadherence as well as from increased rigidity of parasitized red blood cells. No cytoadherence is observed in arterioles of the infected mice despite decreased wall shear rates, indicating that endothelial receptors for cytoadherence are restricted to venules. Infusion of a monoclonal antibody (MAb) against the intercellular adhesion molecule-1 (ICAM-1) resulted in significant increases in both arteriolar and venular Vrbc and wall shear rates, accompanied by detachment of adhered IRBCs at some venular sites. The peripheral blood smears taken after the MAb infusion showed a distinct increase in the percentage of schizonts, again indicating detachment and/or prevention of cytoadherence. An MAb against the vascular cell adhesion molecule-1 (VCAM-1) as well as an irrelevant control antibody had no effect on these parameters. These results provide the first in vivo microcirculatory evidence indicating involvement of ICAM-1, but not of VCAM-1, in the sequestration of IRBCs in a rodent model of cerebral malaria.

Effects of G, a Growth Regulator from Eucalyptus grandis, on Photosynthesis

PubMed Central

Sharkey, Thomas D.; Stevenson, Gay F.; Paton, Dugald M.

1982-01-01

A growth regulator (G; 4-ethyl-1-hydroxy-4,8,8,10,10 pentamethyl-7,9-dioxo-2,3 dioxyabicyclo (4.4.0) decene-5) from Eucalyptus grandis (Maiden) reduced stomatal conductance and also photosynthetic capacity when fed through the transpiration stream of detached leaves. The concentration of G required for this effect was high (10−4 molar), but the amount of G taken up (dose) was below the level which has previously been found in E. grandis leaves. Similar effects were observed in detached leaves of Xanthium strumarium L. though almost 10 times more G was required. G reduced CO2-dependent O2 evolution from isolated cells of X. strumarium. In spinach (Spinacia oleracea L.) chloroplasts, electron transport through photosystem II was reduced by G. It is proposed that G affects stomatal conductance and photosynthesis by reducing photosystem II activity in both the guard cell chloroplasts and mesophyll cell chloroplasts. PMID:16662322

Femtosecond Laser-Assisted Descemetorhexis: A Novel Technique in Descemet Membrane Endothelial Keratoplasty.

PubMed

Pilger, Daniel; von Sonnleithner, Christoph; Bertelmann, Eckart; Joussen, Antonia M; Torun, Necip

2016-10-01

To explore the feasibility of femtosecond laser-assisted descemetorhexis (DR) to facilitate Descemet membrane endothelial keratoplasty (DMEK) surgery. Six pseudophakic patients suffering from Fuchs' endothelial dystrophy underwent femtosecond laser-assisted DMEK surgery. DR was performed using the LenSx femtosecond laser, followed by manual removal of the Descemet membrane. Optical coherence tomography images were used to measure DR parameters. Patients were followed up for 1 month to examine best corrected visual acuity, endothelial cell loss, flap detachment, and structure of the anterior chamber of the eye. The diameter of the DR approximated the intended diameter closely [mean error of 34 μm (0.45%) and 54 μm (0.67%) in the x- and y-diameter, respectively] and did not require manual correction. The median visual acuity increased from 0.4 logMAR (range 0.6-0.4 logMAR) preoperative to 0.2 logMAR (range 0-0.4 logMAR) postoperative. The median endothelial cell loss was 22% (range 7%-34%). No clinically significant flap detachments were noted. All patients had clear corneas after surgery, and no side effects or damage to structures of the anterior chamber were noted. Femtosecond laser-assisted DR is a safe and precise method for facilitating DMEK surgery.

Laser induced photo-detachment of O2 in DC discharge

NASA Astrophysics Data System (ADS)

J, R. LEGORRETA; J, L. PATIÑO; F, B. YOUSIF

2018-07-01

Determination of the negative ion number density of {{{O}}}{{2}}- and {{{O}}}- in a DC discharge of oxygen plasma was made employing Langmuir probe in conjunction with eclipse laser photo-detachment technique. The temporal evolution of the extra electrons resulting from the photo-detachment of {{{O}}}{{2}}- and {{{O}}}- were used to evaluate the negative ion number density. The ratio of {{{O}}}{{2}}- number density to {{{O}}}- varied from 0.03 to 0.22. Number density of both {{{O}}}{{2}}- and {{{O}}}- increased with increasing power and decreased as the pressure was increased. Electron number density was evaluated from the electron energy distribution function (EEDF) using the I–V recorded characteristic curves. Electron temperature between 2 and 2.7 eV were obtained. Influence of the {{{O}}}{{2}}({a}{{1}}{{{Δ }}}{{g}}) metastable state is discussed.

Cellular Migration and Invasion Uncoupled: Increased Migration Is Not an Inexorable Consequence of Epithelial-to-Mesenchymal Transition

PubMed Central

Schaeffer, Daneen; Somarelli, Jason A.; Hanna, Gabi; Palmer, Gregory M.

2014-01-01

Metastatic dissemination requires carcinoma cells to detach from the primary tumor and invade through the basement membrane. To acquire these characteristics, epithelial tumor cells undergo epithelial-to-mesenchymal transitions (EMT), whereby cells lose polarity and E-cadherin-mediated cell-cell adhesion. Post-EMT cells have also been shown, or assumed, to be more migratory; however, there have been contradictory reports on an immortalized human mammary epithelial cell line (HMLE) that underwent EMT. In the context of carcinoma-associated EMT, it is not yet clear whether the change in migration and invasion must be positively correlated during EMT or whether enhanced migration is a necessary consequence of having undergone EMT. Here, we report that pre-EMT rat prostate cancer (PC) and HMLE cells are more migratory than their post-EMT counterparts. To determine a mechanism for increased epithelial cell migration, gene expression analysis was performed and revealed an increase in epidermal growth factor receptor (EGFR) expression in pre-EMT cells. Indeed, inhibition of EGFR in PC epithelial cells slowed migration. Importantly, while post-EMT PC and HMLE cell lines are less migratory, both remain invasive in vitro and, for PC cells, in vivo. Our study demonstrates that enhanced migration is not a phenotypic requirement of EMT, and migration and invasion can be uncoupled during carcinoma-associated EMT. PMID:25002532

Detached Eddy Simulations of Hypersonic Transition

NASA Technical Reports Server (NTRS)

Yoon, S.; Barnhardt, M.; Candler, G.

2010-01-01

This slide presentation reviews the use of Detached Eddy Simulation (DES) of hypersonic transistion. The objective of the study was to investigate the feasibility of using CFD in general, DES in particular, for prediction of roughness-induced boundary layer transition to turbulence and the resulting increase in heat transfer.

Effect of zinc on nectar secretion of Hibiscus rosa -sinensis L.

PubMed

Sawidis, Thomas; Papadopoulou, Alexandra; Voulgaropoulou, Maria

2014-05-01

Zinc toxicity in secretory cells caused a range of effects, mainly depending on metal concentration. Low concentrations activated nectary function increasing nectar secretion but secretion was greatly inhibited or stopped entirely by ongoing concentration. Water loss rate of zinc treated flower parts was significantly reduced whereas green sepals were dehydrated more rapidly in comparison to colored petals. The content of zinc, calcium, magnesium and manganese increased mainly in sepals under excess of zinc, but in the secreted nectar this metal was not evident. Morphological changes were observed in mucilage cells concerning the mucilage structure and appearance. The parenchymatic, subglandular cells displayed an early vacuolarization and cytoplasm condensation. Secretory hairs appeared to be thinner, the apical cell folded inwards and plasmolytic shrinkage became severe in all cells. The waxy cuticula showed an increased electron density. A plasmalemma detachment from the external cell walls was observed creating a gap between cell wall and plasmalemma. ER cisterns of all treated nectary hairs dominated the cytoplasm and electron dense deposits were seen within its profiles. A great number of other organelles were also present, showing electron dense deposits in their membranes as well. The vacuome was drastically reduced in all cells, except in the subglandular ones and electron dense membrane remnants were observed.

Combining light-harvesting with detachability in high-efficiency thin-film silicon solar cells.

PubMed

Ram, Sanjay K; Desta, Derese; Rizzoli, Rita; Bellettato, Michele; Lyckegaard, Folmer; Jensen, Pia B; Jeppesen, Bjarke R; Chevallier, Jacques; Summonte, Caterina; Larsen, Arne Nylandsted; Balling, Peter

2017-06-01

Efforts to realize thin-film solar cells on unconventional substrates face several obstacles in achieving good energy-conversion efficiency and integrating light-management into the solar cell design. In this report a technique to circumvent these obstacles is presented: transferability and an efficient light-harvesting scheme are combined for thin-film silicon solar cells by the incorporation of a NaCl layer. Amorphous silicon solar cells in p-i-n configuration are fabricated on reusable glass substrates coated with an interlayer of NaCl. Subsequently, the solar cells are detached from the substrate by dissolution of the sacrificial NaCl layer in water and then transferred onto a plastic sheet, with a resultant post-transfer efficiency of 9%. The light-trapping effect of the surface nanotextures originating from the NaCl layer on the overlying solar cell is studied theoretically and experimentally. The enhanced light absorption in the solar cells on NaCl-coated substrates leads to significant improvement in the photocurrent and energy-conversion efficiency in solar cells with both 350 and 100 nm thick absorber layers, compared to flat-substrate solar cells. Efficient transferable thin-film solar cells hold a vast potential for widespread deployment of off-grid photovoltaics and cost reduction.

The structural and functional organization of the vestibular apparatus of rats exposed to weightlessness for 20 days on board the Sputnik "Kosmos-782".

PubMed

Vinnikov, Y A; Gazenko, O G; Titova, L K; Bronstein, A A; Govardovskii, V I; Gribakin, F G; Pevzner, R A; Aronova, M Z; Kharkeevich, T A; Tsirulis, T P; Pyatkina, G A; Lichakov, D V; Pal'mbach, L P; Anichin, V F

1979-01-01

This investigation of the vestibular apparatus of rats exposed for 20 days to weightlessness on board an earth satellite and to acceleration during take-off and landing has revealed a set of changes in the structural and functional organization, such as adjoinment of the otolith to the utricle receptor surface and peripheral localization of the nucleoli inside the receptor cells' nuclei. Destruction of some receptor cells, apparently due to increased swelling of the vestibular apparatus tissue and alteration of the shape and structure of the otoconia were observed. In the horizontal crista, detachment of the cupula took place.

Role of macrophage migration inhibitory factor (MIF) in the effects of oxidative stress on human retinal pigment epithelial cells.

PubMed

Ko, Ji-Ae; Sotani, Yasuyuki; Ibrahim, Diah Gemala; Kiuchi, Yoshiaki

2017-10-01

Proliferative vitreoretinopathy (PVR) is the major cause of treatment failure in individuals who undergo surgery for retinal detachment. The epithelial-mesenchymal transition (EMT) in retinal pigment epithelium (RPE) cells contributes to the pathogenesis of PVR. Oxidative stress is thought to play a role in the progression of retinal diseases including PVR. We have now examined the effects of oxidative stress on the EMT and related processes in the human RPE cell line. We found that H 2 O 2 induced the contraction of RPE cells in a three-dimensional collagen gel. Analysis of a cytokine array revealed that H 2 O 2 specifically increased the release of macrophage migration inhibitory factor (MIF) from RPE cells. Reverse transcription-polymerase chain reaction and immunoblot analyses showed that H 2 O 2 increased the expression of MIF in RPE cells. Immunoblot and immunofluorescence analyses revealed that H 2 O 2 upregulated the expression of α-SMA and vimentin and downregulated that of ZO-1 and N-cadherin. Consistent with these observations, the transepithelial electrical resistance of cell was reduced by exposure to H 2 O 2 . The effects of oxidative stress on EMT-related and junctional protein expression as well as on transepithelial electrical resistance were inhibited by antibodies to MIF, but they were not mimicked by treatment with recombinant MIF. Finally, analysis with a profiling array for mitogen-activated protein kinase signalling revealed that H 2 O 2 specifically induced the phosphorylation of p38 mitogen-activated protein kinase. Our results thus suggest that MIF may play a role in induction of the EMT and related processes by oxidative stress in RPE cells and that it might thereby contribute to the pathogenesis of PVR. Proliferative vitreoretinopathy is a major complication of rhegmatogenous retinal detachment, and both oxidative stress and induction of the EMT in RPE cells are thought to contribute to the pathogenesis of this condition. We have now examined the effects of oxidative stress on the EMT and related processes in the human RPE cell line ARPE19. Our results thus implicate MIF in induction of the EMT and related processes by oxidative stress in RPE cells and the regulated expression of EMT markers. They further suggest that MIF may play an important role in the pathogenesis of PVR. Copyright © 2017 John Wiley & Sons, Ltd.

Role for Neutrophil Extracellular Traps (NETs) and Platelet Aggregation in Early Sepsis-induced Hepatic Dysfunction.

PubMed

Sakurai, Kentaro; Miyashita, Tomoharu; Okazaki, Mitsuyoshi; Yamaguchi, Takahisa; Ohbatake, Yoshinao; Nakanuma, Shinichi; Okamoto, Koichi; Sakai, Seisho; Kinoshita, Jun; Makino, Isamu; Nakamura, Keishi; Hayashi, Hironori; Oyama, Katsunobu; Tajima, Hidehiro; Takamura, Hiroyuki; Ninomiya, Itasu; Fushida, Sachio; Harada, Kenichi; Harmon, John W; Ohta, Tetsuo

2017-01-01

Severe sepsis is associated with high morbidity and mortality rates. Inflammation and coagulation play pivotal roles in the pathogenesis of sepsis leading to multiple organ failure, especially in the liver. The aim of the present study was to assess the mechanism from sepsis to liver damage in a mouse model. We created a sepsis model by injecting lipopolysaccharide (LPS) intraperitoneally in mice. At 0, 6, 12, and 24 h following intraperitoneal injection of LPS, mice were euthanised and analyzed. Primary antibodies against myeloperoxidase (MPO), hepatic sinusoidal endothelial cells (SE-1), and P-selectin (CD62p) were used. Expression and localization in neutrophil, sinusoidal endothelial, and platelet cells were assessed by immunohistochemistry. Immunohistochemical analyses revealed a positive staining for MPO, most abundantly in neutrophil granulocytes, within the hepatic sinusoids immediately after injection. Neutrophil extracellular trap (NET)-like structures stained for MPO, indicating the presence of neutrophils undergoing NETosis, were confirmed at 6 h after LPS administration. SE-1 staining for liver sinusoidal endothelial cells was significantly reduced at 12 h post-LPS administration through sinusoidal endothelial injury or detachment. Furthermore, the presence of extravasated platelets was confirmed in the space of Disse at 24 h after LPS administration. Blood sample analyses showed that white blood cell counts and platelet counts decreased gradually, while MPO amounts increased until 12 h after LPS administration. We conclude that NET formation and intravasated platelet aggregation are the first steps from sepsis to liver damage, and that extravasated platelet aggregation promoted by NET-facilitated detachment of sinusoidal endothelial cells is the origin of sepsis-induced liver dysfunction. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

Characterization of edge turbulence in different states of divertor detachment using reflectometry in the ASDEX Upgrade tokamak

NASA Astrophysics Data System (ADS)

Nikolaeva, V.; Guimarais, L.; Manz, P.; Carralero, D.; Manso, M. E.; Stroth, U.; Silva, C.; Conway, G. D.; Seliunin, E.; Vicente, J.; Brida, D.; Aguiam, D.; Santos, J.; Silva, A.; ASDEX Upgrade team; MST1 team

2018-05-01

Transport in the scrape-off layer (SOL) depends on the state of divertor detachment. L-mode discharges were analyzed where the state of divertor detachment is varied through a density ramp-up. By means of reflectometry measurements at the low (LFS) and the high field side (HFS), midplane density fluctuations are studied for the first time in ASDEX Upgrade simultaneously at both sides of the tokamak. Radial density fluctuation profiles (δ {n}e/{n}e) increase with radius in both the HFS and the LFS. It is found that in the SOL density fluctuations at the LFS have about a factor of two larger amplitude than at the HFS in agreement with ballooned transport. Density fluctuations at the LFS show a modest variation with increasing background density resulting mainly from a rise of low frequency components. Experimental results are in good agreement with an enhanced convection of filaments at the LFS at the beginning of outer divertor detachment leading to a flatter SOL density profile. In this phase of the discharge, density fluctuations measured at the HFS far-SOL display a strong increase, which may be associated with the presence of faster filaments originated at the LFS.

[Effect of colcemid on the radial spreading of fibroblasts in culture].

PubMed

Ivanova, O Iu; Komm, S G; Vasil'ev, Iu M; Gel'fand, I M

1977-02-01

Effect of colcemide upon the spreading of mouse embryo fibroblast-like cells on the substrate was studied with the aid of time-lapse microcinematography and scanning electron microscopy. On the glass, colcemide did not prevent the transition of cells into a well-attached state, however, the time needed for this transition was seen considerably increased as compared with the control cultures. Intermediate stages of spreading on flat glass had the following abnormal features in colcemide-containing medium: a) shapes of cytoplasmic outgrowths formed by the cell were altered and their distribution along the cell border appeared less regular; b) partial detachments of the attached parts of cells occurred very frequently; c) the spreading of various parts of the cells was not correlated. Possible mechanisms of colcemide action on the cell spreading are discussed, and it is suggested that intracellular structures sensitive to colcemide are essential for coordination of reactions that occur in various parts of the cell in the course of spreading.

Dynamics of water droplets detached from porous surfaces of relevance to PEM fuel cells.

PubMed

Theodorakakos, A; Ous, T; Gavaises, M; Nouri, J M; Nikolopoulos, N; Yanagihara, H

2006-08-15

The detachment of liquid droplets from porous material surfaces used with proton exchange membrane (PEM) fuel cells under the influence of a cross-flowing air is investigated computationally and experimentally. CCD images taken on a purpose-built transparent fuel cell have revealed that the water produced within the PEM is forming droplets on the surface of the gas-diffusion layer. These droplets are swept away if the velocity of the flowing air is above a critical value for a given droplet size. Static and dynamic contact angle measurements for three different carbon gas-diffusion layer materials obtained inside a transparent air-channel test model have been used as input to the numerical model; the latter is based on a Navier-Stokes equations flow solver incorporating the volume of fluid (VOF) two-phase flow methodology. Variable contact angle values around the gas-liquid-solid contact-line as well as their dynamic change during the droplet shape deformation process, have allowed estimation of the adhesion force between the liquid droplet and the solid surface and successful prediction of the separation line at which droplets loose their contact from the solid surface under the influence of the air stream flowing around them. Parametric studies highlight the relevant importance of various factors affecting the detachment of the liquid droplets from the solid surface.

Stripe-patterned thermo-responsive cell culture dish for cell separation without cell labeling.

PubMed

Kumashiro, Yoshikazu; Ishihara, Jun; Umemoto, Terumasa; Itoga, Kazuyoshi; Kobayashi, Jun; Shimizu, Tatsuya; Yamato, Masayuki; Okano, Teruo

2015-02-11

A stripe-patterned thermo-responsive surface is prepared to enable cell separation without labeling. The thermo-responsive surface containing a 3 μm striped pattern exhibits various cell adhesion and detachment properties. A mixture of three cell types is separated on the patterned surface based on their distinct cell-adhesion properties, and the composition of the cells is analyzed by flow cytometry. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Three-dimensional alginate spheroid culture system of murine osteosarcoma.

PubMed

Akeda, Koji; Nishimura, Akinobu; Satonaka, Haruhiko; Shintani, Ken; Kusuzaki, Katsuyuki; Matsumine, Akihiko; Kasai, Yuichi; Masuda, Koichi; Uchida, Atsumasa

2009-11-01

Osteosarcoma (OS) is the most common primary malignant tumor of the bone and often forms pulmonary metastases, which are the most important prognostic factor. For further elucidation of the mechanism underlying the progression and metastasis of human OS, a culture system mimicking the microenvironment of the tumor in vivo is needed. We report a novel three-dimensional (3D) alginate spheroid culture system of murine osteosarcoma. Two different metastatic clones, the parental Dunn and its derivative line LM8, which has a higher metastatic potential to the lungs, were encapsulated in alginate beads to develop the 3D culture system. The beads containing murine OS cells were also transplanted into mice to determine their metastatic potential in vivo. In this culture system, murine OS cells encapsulated in alginate beads were able to grow in a 3D structure with cells detaching from the alginate environment. The number of detaching cells was higher in the LM8 cell line than the Dunn cell line. In the in vivo alginate bead transplantation model, the rate of pulmonary metastasis was higher with LM8 cells compared with that of Dunn cells. The cell characteristics and kinetics in this culture system closely reflect the original malignant potential of the cells in vivo.

Role of biofilm roughness and hydrodynamic conditions in Legionella pneumophila adhesion to and detachment from simulated drinking water biofilms.

PubMed

Shen, Yun; Monroy, Guillermo L; Derlon, Nicolas; Janjaroen, Dao; Huang, Conghui; Morgenroth, Eberhard; Boppart, Stephen A; Ashbolt, Nicholas J; Liu, Wen-Tso; Nguyen, Thanh H

2015-04-07

Biofilms in drinking water distribution systems (DWDS) could exacerbate the persistence and associated risks of pathogenic Legionella pneumophila (L. pneumophila), thus raising human health concerns. However, mechanisms controlling adhesion and subsequent detachment of L. pneumophila associated with biofilms remain unclear. We determined the connection between L. pneumophila adhesion and subsequent detachment with biofilm physical structure characterization using optical coherence tomography (OCT) imaging technique. Analysis of the OCT images of multispecies biofilms grown under low nutrient condition up to 34 weeks revealed the lack of biofilm deformation even when these biofilms were exposed to flow velocity of 0.7 m/s, typical flow for DWDS. L. pneumophila adhesion on these biofilm under low flow velocity (0.007 m/s) positively correlated with biofilm roughness due to enlarged biofilm surface area and local flow conditions created by roughness asperities. The preadhered L. pneumophila on selected rough and smooth biofilms were found to detach when these biofilms were subjected to higher flow velocity. At the flow velocity of 0.1 and 0.3 m/s, the ratio of detached cell from the smooth biofilm surface was from 1.3 to 1.4 times higher than that from the rough biofilm surface, presumably because of the low shear stress zones near roughness asperities. This study determined that physical structure and local hydrodynamics control L. pneumophila adhesion to and detachment from simulated drinking water biofilm, thus it is the first step toward reducing the risk of L. pneumophila exposure and subsequent infections.

Retinal Detachment Associated with AIDS-Related Cytomegalovirus Retinitis: Risk Factors in a Resource-Limited Setting

PubMed Central

Yen, Michael; Chen, Jenny; Ausayakhun, Somsanguan; Kunavisarut, Paradee; Vichitvejpaisal, Pornpattana; Ausayakhun, Sakarin; Jirawison, Choeng; Shantha, Jessica; Holland, Gary N; Heiden, David; Margolis, Todd P; Keenan, Jeremy D

2014-01-01

Purpose To determine risk factors predictive of retinal detachment in patients with cytomegalovirus (CMV) retinitis in a setting with limited access to ophthalmic care. Design Case-control study. Methods Sixty-four patients with CMV retinitis and retinal detachment were identified from the Ocular Infectious Diseases and Retina Clinics at Chiang Mai University. Three control patients with CMV retinitis but no retinal detachment were selected for each case, matched by calendar date. The medical records of each patient were reviewed, with patient-level and eye-level features recorded for the clinic visit used to match cases and controls, and also for the initial clinic visit at which CMV retinitis was diagnosed. Risk factors for retinal detachment were assessed separately for each of these time points using multivariate conditional logistic regression models that included 1 eye from each patient. Results Patients with a retinal detachment were more likely than controls to have low visual acuity (OR, 1.24 per line of worse vision on the logMAR scale; 95%CI, 1.16-1.33) and bilateral disease (OR, 2.12; 95%CI, 0.92-4.90). Features present at the time of the initial diagnosis of CMV retinitis that predicted subsequent retinal detachment included bilateral disease (OR, 2.68; 95%CI, 1.18-6.08) and lesion size (OR, 2.64 per 10% increase in lesion size; 95%CI, 1.41-4.94). Conclusion Bilateral CMV retinitis and larger lesion sizes, each of which is a marker of advanced disease, were associated with subsequent retinal detachment. Earlier detection and treatment may reduce the likelihood that patients with CMV retinitis develop a retinal detachment. PMID:25448999

The Detached Auroras Induced by the Solar Wind Pressure Enhancement in Both Hemispheres From Imaging and In Situ Particle Observations

NASA Astrophysics Data System (ADS)

Zhou, Su; Luan, Xiaoli; Søraas, Finn; Østgaard, Nikolai; Raita, Tero

2018-04-01

This paper presents simultaneous detached proton auroras that appeared in both hemispheres at 11:06 UT, 08 March 2012, just 2 min after a sudden solar wind pressure enhancement ( 11:04 UT) hit the Earth. They were observed under northward interplanetary magnetic field Bz condition and during the recovery phase of a moderate geomagnetic storm. In the Northern Hemisphere, Defense Meteorological Satellite Program/Special Sensor Ultraviolet Spectrographic Imager observed that the detached arc occurred within 60°-65° magnetic latitude and covered a few magnetic local time (MLT) hours ranging from 0530 to 0830 MLT with a possible extension toward noon. At the same time (11:06 UT), Polar Orbiting Environment Satellites 19 detected a detached proton aurora around 1300 MLT in the Southern Hemisphere, centering 62° magnetic latitude, which was at the same latitudes as the northern detached arc. This southern aurora was most probably a part of a dayside detached arc that was conjugate to the northern one. In situ particle observations indicated that the detached auroras were dominated by protons/ions with energies ranging from around 20 keV to several hundreds of keV, without obvious electron precipitations. These detached arcs persisted for less than 6 min, consistent with the impact from pressure enhancement and the observed electromagnetic ion cyclotron (EMIC) waves. It is suggested that the increasing solar wind pressure pushed the hot ions in the ring current closer to Earth where the steep gradient of cold plasma favored EMIC wave growth. By losing energy to EMIC waves the energetic protons (>20 keV) were scattered into the loss cone and produced the observed detached proton auroras.

Retinal detachment associated with AIDS-related cytomegalovirus retinitis: risk factors in a resource-limited setting.

PubMed

Yen, Michael; Chen, Jenny; Ausayakhun, Somsanguan; Kunavisarut, Paradee; Vichitvejpaisal, Pornpattana; Ausayakhun, Sakarin; Jirawison, Choeng; Shantha, Jessica; Holland, Gary N; Heiden, David; Margolis, Todd P; Keenan, Jeremy D

2015-01-01

To determine risk factors predictive of retinal detachment in patients with cytomegalovirus (CMV) retinitis in a setting with limited access to ophthalmic care. Case-control study. Sixty-four patients with CMV retinitis and retinal detachment were identified from the Ocular Infectious Diseases and Retina Clinics at Chiang Mai University. Three control patients with CMV retinitis but no retinal detachment were selected for each case, matched by calendar date. The medical records of each patient were reviewed, with patient-level and eye-level features recorded for the clinic visit used to match cases and controls, and also for the initial clinic visit at which CMV retinitis was diagnosed. Risk factors for retinal detachment were assessed separately for each of these time points using multivariate conditional logistic regression models that included 1 eye from each patient. Patients with a retinal detachment were more likely than controls to have low visual acuity (odds ratio [OR], 1.24 per line of worse vision on the logMAR scale; 95% confidence interval [CI], 1.16-1.33) and bilateral disease (OR, 2.12; 95% CI, 0.92-4.90). Features present at the time of the initial diagnosis of CMV retinitis that predicted subsequent retinal detachment included bilateral disease (OR, 2.68; 95% CI, 1.18-6.08) and lesion size (OR, 2.64 per 10% increase in lesion size; 95% CI, 1.41-4.94). Bilateral CMV retinitis and larger lesion sizes, each of which is a marker of advanced disease, were associated with subsequent retinal detachment. Earlier detection and treatment may reduce the likelihood that patients with CMV retinitis develop a retinal detachment. Copyright © 2015 Elsevier Inc. All rights reserved.

Miniature Dielectric Barrier Discharge Nonthermal Plasma Induces Apoptosis in Lung Cancer Cells and Inhibits Cell Migration.

PubMed

Karki, Surya B; Yildirim-Ayan, Eda; Eisenmann, Kathryn M; Ayan, Halim

2017-01-01

Traditional cancer treatments like radiotherapy and chemotherapy have drawbacks and are not selective for killing only cancer cells. Nonthermal atmospheric pressure plasmas with dielectric barrier discharge (DBD) can be applied to living cells and tissues and have emerged as novel tools for localized cancer therapy. The purpose of this study was to investigate the different effects caused by miniature DBD (mDBD) plasma to A549 lung cancer cells. In this study, A549 lung cancer cells cultured in 12 well plates were treated with mDBD plasma for specified treatment times to assess the changes in the size of the area of cell detachment, the viability of attached or detached cells, and cell migration. Furthermore, we investigated an innovative mDBD plasma-based therapy for localized treatment of lung cancer cells through apoptotic induction. Our results indicate that plasma treatment for 120 sec causes apoptotic cell death in 35.8% of cells, while mDBD plasma treatment for 60 sec, 30 sec, or 15 sec causes apoptotic cell death in 20.5%, 14.1%, and 6.3% of the cell population, respectively. Additionally, we observed reduced A549 cell migration in response to mDBD plasma treatment. Thus, mDBD plasma system can be a viable platform for localized lung cancer therapy.

Onset of vitellogenin production and vitellogenesis, and their relationship to changes in the midgut epithelium and oocytes in the tick Dermacentor variabilis.

PubMed

Coons, L B; Lamoreaux, W J; Rosell-Davis, R; Tarnowski, B I

1989-05-01

In Dermacentor variabilis (Say), the onset of vitellogenin production and vitellogenesis (uptake of vitellogenin into oocytes) began during the rapid-engorgement feeding period. Mating was required for both vitellogenin production and vitellogenesis to complete the tick's life cycle. Complete immunological identity, as measured by Ouchterlony's double diffusion test, existed between vitellogenin from the fat body, midgut and hemolymph, and vitellin from the ovaries and eggs. Antivitellin antibody did not react with host hemoglobin nor with fat body, midgut, and ovary extracts from feeding females prior to rapid engorgement, feeding unmated females, or unfed or fed males. Some unmated females fed for 13 days and then hand-detached from the host eventually began oviposition after going through a preoviposition period. In these ticks, organ extracts from the midgut, fat body and ovary reacted with antivitellin antibody. The presence or absence of presumed vitellogenic cells in the midgut and yolk bodies in oocytes corresponded with the presence or absence of vitellogenin and vitellogenesis as measured by Ouchterlony's test. Presumed vitellogenic cells increased in size during the preoviposition period. These cells reached their greatest size during the time when the most eggs were being produced, and then declined in size toward the end of oviposition. Vitellogenin was deposited directly into developing yolk bodies in oocytes and was not processed through lysosomes. Feeding was the process that initiated the formation of eggshell cuticle. Detachment from the host was required for the initiation of oviposition.

Occupational Lifting Tasks and Retinal Detachment in Non-Myopics and Myopics: Extended Analysis of a Case-Control Study

PubMed Central

Mattioli, Stefano; Curti, Stefania; De Fazio, Rocco; Mt Cooke, Robin; Zanardi, Francesca; Bonfiglioli, Roberta; Violante, Francesco S

2012-01-01

Objectives Lifting heavy weights involves the Valsalva manoeuvre, which leads to intraocular pressure spikes. We used data from a case-control study to further investigate the hypothesis that occupational lifting is a risk factor for retinal detachment. Methods The study population included 48 cases (patients operated for retinal detachment) and 84 controls (outpatients attending an eye clinic). The odds ratios (OR) of idiopathic retinal detachment were estimated with a logistic regression model (adjusted for age, sex and body mass index). Three indexes were used to examine exposure to lifting; 1) maximum load lifted, 2) average weekly lifting, 3) lifelong cumulative lifting. Results For all indexes, the most exposed subjects showed an increased risk of retinal detachment compared with the unexposed (index 1: OR 3.57, 95% confidence interval [CI] 1.21-10.48; index 2: OR 3.24, 95% CI 1.32-7.97; index 3: OR 2.23, 95% CI 1.27-8.74) and dose-response relationships were apparent. Conclusion These results reinforce the hypothesis that heavy occupational lifting may be a relevant risk factor for retinal detachment. PMID:22953231

Modeling of combined effects of divertor closure and advanced magnetic configuration on detachment in DIII-D by SOLPS

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

Si, Hang; Guo, Houyang Y.; Covele, Brent

One of the major challenges facing the design and operation of next-step high-power steady-state fusion devices is to develop a divertor solution for handling power exhaust, while ensuring acceptable divertor target plate erosion, which necessitates access to divertor detachment at relative low main plasma densities compatible with current drive and high plasma confinement. Detailed modeling with SOLPS is carried out to examine the effect of divertor closure on detachment with the normal single null divertor (SD) configuration, as well as one of the advanced divertor configurations, such as x-divertor (XD) respectively. The SOLPS modeling for a high confinement plasma in DIII-D finds that increasing divertor closure with SD reduces the upstream separatrix density at the onset of detachment frommore » $$1.18\\times {{10}^{19}}\\,{{{\\rm m}}^{-3}}$$ to $$0.88\\times {{10}^{19}}\\,{{{\\rm m}}^{-3}}$$. Furthermore, coupling the divertor closure with XD further promotes the onset of divertor detachment at a still lower upstream separatrix density, down to the value of $$0.67\\times {{10}^{19}}\\,{{{\\rm m}}^{-3}}$$, thus, showing that divertor closure and advanced magnetic configuration can work synergistically to facilitate divertor detachment.« less

Modeling of combined effects of divertor closure and advanced magnetic configuration on detachment in DIII-D by SOLPS

DOE PAGES

Si, Hang; Guo, Houyang Y.; Covele, Brent; ...

2018-04-04

One of the major challenges facing the design and operation of next-step high-power steady-state fusion devices is to develop a divertor solution for handling power exhaust, while ensuring acceptable divertor target plate erosion, which necessitates access to divertor detachment at relative low main plasma densities compatible with current drive and high plasma confinement. Detailed modeling with SOLPS is carried out to examine the effect of divertor closure on detachment with the normal single null divertor (SD) configuration, as well as one of the advanced divertor configurations, such as x-divertor (XD) respectively. The SOLPS modeling for a high confinement plasma in DIII-D finds that increasing divertor closure with SD reduces the upstream separatrix density at the onset of detachment frommore » $$1.18\\times {{10}^{19}}\\,{{{\\rm m}}^{-3}}$$ to $$0.88\\times {{10}^{19}}\\,{{{\\rm m}}^{-3}}$$. Furthermore, coupling the divertor closure with XD further promotes the onset of divertor detachment at a still lower upstream separatrix density, down to the value of $$0.67\\times {{10}^{19}}\\,{{{\\rm m}}^{-3}}$$, thus, showing that divertor closure and advanced magnetic configuration can work synergistically to facilitate divertor detachment.« less

Detachment experiments in new DIII-D upper divertor

NASA Astrophysics Data System (ADS)

Moser, A. L.; Leonard, A. W.; Groebner, R. J.; Guo, H.; Wang, H.; Watkins, J. G.; McLean, A. G.; Fenstermacher, M. E.; Shafer, M. W.; Briesemeister, A. R.; Hinson, E. T.

2017-10-01

Installation of the Small Angle Slot (SAS) in the upper divertor of DIII-D enables new studies of the effect of target and baffle geometry on divertor detachment. This structure provides a more-closed upper divertor as well as the SAS divertor itself. Initial SAS experiment results indicate that divertor detachment occurs at a lower line-averaged density than in the more-open, lower single null divertor configurations on DIII-D. In contrast, the increased divertor closure of the new installation did not reduce the upstream density required for detachment beyond that achieved with the previous upper divertor structure. Particle pumping in the upper divertor structure is found to produce a 10 % reduction in the pedestal density required for detachment compared to the case with no pumping. Comparisons focus on both the onset of detachment (measured by in-target Langmuir probes) as a function of upstream density, as well as the effect of the new divertor configurations on pedestal density profiles. Work supported by US DOE under DE-FC02-04ER54698, DE-AC05-00OR22725, DE-AC04-94AL85000, DE-AC52-07NA27344, and DE-SC00013911.

Modeling of combined effects of divertor closure and advanced magnetic configuration on detachment in DIII-D by SOLPS

NASA Astrophysics Data System (ADS)

Si, H.; Guo, H. Y.; Covele, B.; Leonard, A. W.; Watkins, J. G.; Thomas, D.; Ding, R.

2018-05-01

One of the major challenges facing the design and operation of next-step high-power steady-state fusion devices is to develop a divertor solution for handling power exhaust, while ensuring acceptable divertor target plate erosion, which necessitates access to divertor detachment at relative low main plasma densities compatible with current drive and high plasma confinement. Detailed modeling with SOLPS is carried out to examine the effect of divertor closure on detachment with the normal single null divertor (SD) configuration, as well as one of the advanced divertor configurations, such as x-divertor (XD) respectively. The SOLPS modeling for a high confinement plasma in DIII-D finds that increasing divertor closure with SD reduces the upstream separatrix density at the onset of detachment from 1.18× {{10}19} {{m}-3} to 0.88× {{10}19} {{m}-3} . Moreover, coupling the divertor closure with XD further promotes the onset of divertor detachment at a still lower upstream separatrix density, down to the value of 0.67× {{10}19} {{m}-3} , thus, showing that divertor closure and advanced magnetic configuration can work synergistically to facilitate divertor detachment.

Transient adhesion in a non-fully detached contact.

PubMed

Liu, Zheyu; Lu, Hongyu; Zheng, Yelong; Tao, Dashuai; Meng, Yonggang; Tian, Yu

2018-04-18

Continuous approaching and detaching displacement usually occurs in an adhesion test. Here, we found a transient adhesion force at the end of a non-fully detached contact. This force occurred when the nominal detaching displacement was less than the traditional quasi-static theory predicted zero force point. The transient adhesion force was ascribed to interfacial adhesion hysteresis, which was caused by the cracking process of the contact and the deformation competition between the sphere and supporting spring. Results indicated that the testing of adhesion can be significantly affected by different combinations of stiffnesses of the contact objects and the supporting spring cantilever. This combination should be carefully designed in an adhesion test. All these results enabled increased understanding of the nature of adhesion and can guide the design of adhesive actuators.

Standardization of the Descemet membrane endothelial keratoplasty technique: Outcomes of the first 450 consecutive cases.

PubMed

Satué, M; Rodríguez-Calvo-de-Mora, M; Naveiras, M; Cabrerizo, J; Dapena, I; Melles, G R J

2015-08-01

To evaluate the clinical outcome of the first 450 consecutive cases after Descemet membrane endothelial keratoplasty (DMEK), as well as the effect of standardization of the technique. Comparison between 3 groups: Group I: (cases 1-125), as the extended learning curve; Group II: (cases 126-250), transition to technique standardization; Group III: (cases 251-450), surgery with standardized technique. Best corrected visual acuity, endothelial cell density, pachymetry and intra- and postoperative complications were evaluated before, and 1, 3 and 6 months after DMEK. At 6 months after surgery, 79% of eyes reached a best corrected visual acuity of≥0.8 and 43%≥1.0. Mean preoperative endothelial cell density was 2,530±220 cells/mm2 and 1,613±495 at 6 months after surgery. Mean pachymetry measured 668±92 μm and 526±46 μm pre- and (6 months) postoperatively, respectively. There were no significant differences in best corrected visual acuity, endothelial cell density and pachymetry between the 3 groups (P > .05). Graft detachment presented in 17.3% of the eyes. The detachment rate declined from 24% to 12%, and the rate of secondary surgeries from 9.6% to 3.5%, from group I to III respectively. Visual outcomes and endothelial cell density after DMEK are independent of the technique standardization. However, technique standardization may have contributed to a lower graft detachment rate and a relatively low number of secondary interventions required. As such, DMEK may become the first choice of treatment in corneal endothelial disease. Copyright © 2014 Sociedad Española de Oftalmología. Published by Elsevier España, S.L.U. All rights reserved.

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

Ahn, J. -W.; Briesemester, A. R.; Kobayashi, M.

Enhanced perpendicular heat and momentum transport induces parallel pressure loss leading to divertor detachment, which can be produced by the increase of density in 2D tokamaks. However, in the 3D configurations such as tokamaks with 3D fields and stellarators, the fraction of perpendicular transport can be higher even in a lower density regime, which could lead to the early transition to detachment without passing through the high-recycling regime. 3D fields applied to the limiter tokamak plasmas produce edge stochastic layers close to the last closed flux surface (LCFS), which can allow for enhanced perpendicular transport and indeed the absence ofmore » high recycling regime and early detachment have been observed in TEXTOR and Tore Supra. However, in the X-point divertor tokamaks with the applied 3D fields, the parallel transport is still dominant and the detachment facilitation has not been observed yet. Rather, 3D fields affected detachment adversely under certain conditions, either by preventing detachment onset as seen in DIII-D or by re-attaching the existing detached plasma as shown in NSTX. The possible way for strong 3D effects to induce access to the early detachment in divertor tokamaks appears to be via significant perpendicular loss of parallel momentum by frictional force for the counter-streaming flows between neighboring flow channels in the divertor. In principle, the adjacent lobes in the 3D divertor tokamak may generate the counter-streaming flow channels. However, an EMC3-EIRENE simulation for ITER H-mode plasmas demonstrated that screened RMP leads to significantly reduced counter-flows near the divertor target, therefore the momentum loss effect leading to detachment facilitation is expected to be small. This is consistent with the observation in LHD, which showed screening (amplification) of RMP fields in the attachment (stable detachment) case. In conclusion, work for optimal parameter window for best divertor operation scenario is needed particularly for the 3D divertor tokamak configuration.« less

Growth and detachment of single hydrogen bubbles in a magnetohydrodynamic shear flow

NASA Astrophysics Data System (ADS)

Baczyzmalski, Dominik; Karnbach, Franziska; Mutschke, Gerd; Yang, Xuegeng; Eckert, Kerstin; Uhlemann, Margitta; Cierpka, Christian

2017-09-01

This study investigates the effect of a magnetohydrodynamic (MHD) shear flow on the growth and detachment of single sub-millimeter-sized hydrogen gas bubbles. These bubbles were electrolytically generated at a horizontal Pt microelectrode (100 μ m in diameter) in an acidic environment (1 M H2SO4 ). The inherent electric field was superimposed by a homogeneous electrode-parallel magnetic field of up to 700 mT to generate Lorentz forces in the electrolyte, which drive the MHD flow. The growth and motion of the hydrogen bubble was analyzed by microscopic high-speed imaging and measurements of the electric current, while particle tracking velocimetry (μ PTV ) and particle image velocimetry (μ PIV ) were applied to measure the surrounding electrolyte flow. In addition, numerical flow simulations were performed based on the experimental conditions. The results show a significant reduction of the bubble growth time and detachment diameter with increasing magnetic induction, which is known to improve the efficiency of water electrolysis. In order to gain further insight into the bubble detachment mechanism, an analysis of the forces acting on the bubble was performed. The strong MHD-induced drag force causes the bubble to slowly slide away from the center of the microelectrode before its detachment. This motion increases the active electrode area and enhances the bubble growth rate. The results further indicate that at large current densities the coalescence of tiny bubbles formed at the foot of the main bubble might play an important role for the bubble detachment. Moreover, the occurrence of Marangoni stresses at the gas-liquid interface is discussed.

Freezing pattern and frost killing temperature of apple (Malus domestica) wood under controlled conditions and in nature.

PubMed

Pramsohler, Manuel; Hacker, Jürgen; Neuner, Gilbert

2012-07-01

The freezing pattern and frost killing temperatures of apple (Malus domestica Borkh.) xylem were determined by differential thermal analysis and infrared differential thermal analysis (IDTA). Results from detached or attached twigs in controlled freezing experiments and during natural field freezing of trees were compared. Non-lethal freezing of apoplastic water in apple xylem as monitored during natural winter frosts in the field occurred at -1.9 ± 0.4 °C and did not change seasonally. The pattern of whole tree freezing was variable and specific to the environmental conditions. On detached twigs high-temperature freezing exotherms (HTEs) occurred 2.8 K below the temperature observed under natural frosts in the field with a seasonal mean of -4.7 ± 0.5 °C. Microporous apple xylem showed freezing without a specific pattern within a few seconds in IDTA images during HTEs, which is in contrast to macroporous xylem where a 2D freezing pattern mirrors anatomical structures. The pith tissue always remained unfrozen. Increasing twig length increased ice nucleation temperature; for increased twig diameter the effect was not significant. In attached twigs frozen in field portable freezing chambers, HTEs were recorded at a similar mean temperature (-4.6 ± 1.0 °C) to those for detached twigs. Upon lethal intracellular freezing of apple xylem parenchyma cells (XPCs) low-temperature freezing exotherms (LTEs) can be recorded. Low-temperature freezing exotherms determined on detached twigs varied significantly between a winter minimum of -36.9 °C and a summer maximum -12.7 °C. Within the temperature range wherein LTEs were recorded by IDTA in summer (-12.7 ± 0.5 to -20.3 ± 1.1 °C) various tiny clearly separated discontinuous freezing events could be detected similar to that in other species with contrasting XPC anatomy. These freezing events appeared to be initially located in the primary and only later in the secondary xylem. During the LTE no freezing events in the bark and central pith tissue were recorded. Attached twigs were exposed to various freezing temperatures at which LTEs occur. Even if 60% of XPCs were frost-damaged twigs were able to recuperate and showed full re-growth indicating a high regeneration capacity even after severe frost damage to XPCs.

Adhesion strength of a living cell to various substrates measured using a cup-attached atomic force microscopy chip

NASA Astrophysics Data System (ADS)

Kim, Hyonchol; Ishibashi, Kenta; Matsuo, Kosuke; Kira, Atsushi; Onomura, Yui; Okada, Tomoko; Nakamura, Chikashi

2018-03-01

Cell adhesion strengths to various substrates were quantitatively measured using atomic force microscopy (AFM). A cup-shaped metal hemisphere was attached to the apex of the AFM cantilever, the “cup-chip” approached a cell (FP10SC2) to pick it up, the captured cell approached any one of six different substrates [gold (Au), nickel (Ni), bovine serum albumin (BSA), an amino group (NH2), poly(tetrafluoroethylene) (PTFE), and structured PTFE (sPTFE)], and the cell adhesion strength at the initial contact period was evaluated by detaching the cell from the substrate. The results obtained showed that the force needed to detach the cell from the NH2 substrate was more than 3-fold larger than that of metal substrates (Au and Ni), more than 15-fold larger than that of biochemically treated substrates (BSA), and more than 20-fold larger than that of hydrophobic substrates (PTFE and sPTFE). Using differences in adhesion strengths, a cell on a sPTFE substrate was picked up using a BSA-coated cup-chip, placed on a NH2 substrate, repeating this cell manipulation five times, and line patterning of cells was achieved. These results indicate that measurements of cell adhesion strength are fundamental to fabricate desired cell networks and the cup-chip is a useful tool for achieving easy cell manipulation.

Proapoptotic protein Bim attenuates estrogen-enhanced survival in lymphangioleiomyomatosis

PubMed Central

Li, Chenggang; Li, Na; Liu, Xiaolei; Zhang, Erik Y.; Sun, Yang; Masuda, Kouhei; Li, Jing; Sun, Julia; Morrison, Tasha; Li, Xiangke; Chen, Yuanguang; Wang, Jiang; Karim, Nagla A.; Zhang, Yi; Blenis, John; Reginato, Mauricio J.; Henske, Elizabeth P.; Yu, Jane J.

2016-01-01

Lymphangioleiomyomatosis (LAM) is a progressive lung disease that primarily affects young women. Genetic evidence suggests that LAM cells bearing TSC2 mutations migrate to the lungs, proliferate, and cause cystic remodeling. The female predominance indicates that estrogen plays a critical role in LAM pathogenesis, and we have proposed that estrogen promotes LAM cell metastasis by inhibition of anoikis. We report here that estrogen increased LAM patient–derived cells’ resistance to anoikis in vitro, accompanied by decreased accumulation of the proapoptotic protein Bim, an activator of anoikis. The resistance to anoikis was reversed by the proteasome inhibitor, bortezomib. Treatment of LAM patient–derived cells with estrogen plus bortezomib promoted anoikis compared with estrogen alone. Depletion of Bim by siRNA in TSC2-deficient cells resulted in anoikis resistance. Treatment of mice with bortezomib reduced estrogen-promoted lung colonization of TSC2-deficient cells. Importantly, molecular depletion of Bim by siRNA in Tsc2-deficient cells increased lung colonization in a mouse model. Collectively, these data indicate that Bim plays a key role in estrogen-enhanced survival of LAM patient–derived cells under detached conditions that occur with dissemination. Thus, targeting Bim may be a plausible future treatment strategy in patients with LAM. PMID:27882343

Variations in soil detachment rates after wildfire as a function of soil depth, flow properties, and root properties

USGS Publications Warehouse

Moody, John A.; Nyman, Peter

2013-01-01

Wildfire affects hillslope erosion through increased surface runoff and increased sediment availability, both of which contribute to large post-fire erosion events. Relations between soil detachment rate, soil depth, flow and root properties, and fire impacts are poorly understood and not represented explicitly in commonly used post-fire erosion models. Detachment rates were measured on intact soil cores using a modified tilting flume. The cores were mounted flush with the flume-bed and a measurement was made on the surface of the core. The core was extruded upward, cut off, and another measurement was repeated at a different depth below the original surface of the core. Intact cores were collected from one site burned by the 2010 Fourmile Canyon (FMC) fire in Colorado and from one site burned by the 2010 Pozo fire in California. Each site contained contrasting vegetation and soil types. Additional soil samples were collected alongside the intact cores and were analyzed in the laboratory for soil properties (organic matter, bulk density, particle-size distribution) and for root properties (root density and root-length density). Particle-size distribution and root properties were different between sites, but sites were similar in terms of bulk density and organic matter. Soil detachment rates had similar relations with non-uniform shear stress and non-uniform unit stream power. Detachment rates within single sampling units displayed a relatively weak and inconsistent relation to flow variables. When averaged across all clusters, the detachment rate displayed a linear relation to shear stress, but variability in soil properties meant that the shear stress accounted for only a small proportion of the overall variability in detachment rates (R2 = 0.23; R2 is the coefficient of determination). Detachment rate was related to root-length density in some clusters (R2 values up to 0.91) and unrelated in others (R2 values 2 value improved and the range of exponents became narrower by applying a multivariate regression model where boundary shear stress and root-length density were included as explanatory variables. This suggests that an erodibility parameter which incorporates the effects of both flow and root properties on detachment could improve the representation of sediment availability after wildfire.

Impact of 10% SF6 Gas Compared to 100% Air Tamponade in Descemet's Membrane Endothelial Keratoplasty.

PubMed

Rickmann, Annekatrin; Szurman, Peter; Jung, Sacha; Boden, Karl Thomas; Wahl, Silke; Haus, Arno; Boden, Katrin; Januschowski, Kai

2018-04-01

To compare the clinical outcomes following Descemet's membrane endothelial keratoplasty (DMEK) with 100% air tamponade versus 10% sulfur hexafluoride (SF 6 ) tamponade. Retrospective analysis of 108 consecutive DMEK cases subdivided by anterior chamber tamponade with 54 eyes receiving 10% SF 6 and 54 eyes receiving 100% air injection. A post-hoc matched analysis revealed no statistically significant differences between the groups. The main outcome measurements were the complication rate, including intra- and postoperative complications and graft detachment rate requiring re-bubbling. Clinical outcome included best-corrected visual acuity (BCVA), endothelial cell count (ECC), and central corneal thickness (CCT) measured 1, 3, and 6 months after DMEK surgery. The graft detachment rate with consecutive re-bubbling was 18.5% in the air group and 22.2% in the SF 6 group (p = 0.2). Remaining small peripheral graft detachments with a clear cornea occurred more often in the 100% air group (air: 22.2%; 12/54, 6/12 inferior compared to SF 6 : 7.4%; 4/54, 2/4 inferior; p = 0.06). The primary graft failure rate was comparable between the two groups. No complete graft detachment occurred. Outcome results for BCVA, ECC, and CCT at all follow-up time points were comparable between the two groups. The clinical outcomes (including re-bubbling rate, primary graft failure rate, and endothelial cell loss) were comparable with 100% air versus 10% SF 6 tamponade, whereas other studies suggest that a higher SF 6 concentration (20%) may result in a lower re-bubbling rate.

Glycolysis is the primary bioenergetic pathway for cell motility and cytoskeletal remodeling in human prostate and breast cancer cells.

PubMed

Shiraishi, Takumi; Verdone, James E; Huang, Jessie; Kahlert, Ulf D; Hernandez, James R; Torga, Gonzalo; Zarif, Jelani C; Epstein, Tamir; Gatenby, Robert; McCartney, Annemarie; Elisseeff, Jennifer H; Mooney, Steven M; An, Steven S; Pienta, Kenneth J

2015-01-01

The ability of a cancer cell to detach from the primary tumor and move to distant sites is fundamental to a lethal cancer phenotype. Metabolic transformations are associated with highly motile aggressive cellular phenotypes in tumor progression. Here, we report that cancer cell motility requires increased utilization of the glycolytic pathway. Mesenchymal cancer cells exhibited higher aerobic glycolysis compared to epithelial cancer cells while no significant change was observed in mitochondrial ATP production rate. Higher glycolysis was associated with increased rates of cytoskeletal remodeling, greater cell traction forces and faster cell migration, all of which were blocked by inhibition of glycolysis, but not by inhibition of mitochondrial ATP synthesis. Thus, our results demonstrate that cancer cell motility and cytoskeleton rearrangement is energetically dependent on aerobic glycolysis and not oxidative phosphorylation. Mitochondrial derived ATP is insufficient to compensate for inhibition of the glycolytic pathway with regard to cellular motility and CSK rearrangement, implying that localization of ATP derived from glycolytic enzymes near sites of active CSK rearrangement is more important for cell motility than total cellular ATP production rate. These results extend our understanding of cancer cell metabolism, potentially providing a target metabolic pathway associated with aggressive disease.

Glycolysis is the primary bioenergetic pathway for cell motility and cytoskeletal remodeling in human prostate and breast cancer cells

PubMed Central

Shiraishi, Takumi; Verdone, James E.; Huang, Jessie; Kahlert, Ulf D.; Hernandez, James R.; Torga, Gonzalo; Zarif, Jelani C.; Epstein, Tamir; Gatenby, Robert; McCartney, Annemarie; Elisseeff, Jennifer H.; Mooney, Steven M.; An, Steven S.; Pienta, Kenneth J.

2015-01-01

The ability of a cancer cell to detach from the primary tumor and move to distant sites is fundamental to a lethal cancer phenotype. Metabolic transformations are associated with highly motile aggressive cellular phenotypes in tumor progression. Here, we report that cancer cell motility requires increased utilization of the glycolytic pathway. Mesenchymal cancer cells exhibited higher aerobic glycolysis compared to epithelial cancer cells while no significant change was observed in mitochondrial ATP production rate. Higher glycolysis was associated with increased rates of cytoskeletal remodeling, greater cell traction forces and faster cell migration, all of which were blocked by inhibition of glycolysis, but not by inhibition of mitochondrial ATP synthesis. Thus, our results demonstrate that cancer cell motility and cytoskeleton rearrangement is energetically dependent on aerobic glycolysis and not oxidative phosphorylation. Mitochondrial derived ATP is insufficient to compensate for inhibition of the glycolytic pathway with regard to cellular motility and CSK rearrangement, implying that localization of ATP derived from glycolytic enzymes near sites of active CSK rearrangement is more important for cell motility than total cellular ATP production rate. These results extend our understanding of cancer cell metabolism, potentially providing a target metabolic pathway associated with aggressive disease. PMID:25426557

Multi-scale cell/surface interaction on modified titanium aluminum vanadium surfaces

NASA Astrophysics Data System (ADS)

Chen, Jianbo

This dissertation presents a series of experimental studies of the effects of multi-scale cell/surface interactions on modified Ti-6Al-4V surfaces. These include laser-grooved surfaces; porous structures and RGD-coated laser-grooved surfaces. A nano-second DPSS UV lasers with a Gaussian pulse energy profile was used to introduce the desired micro-groove geometries onto Ti-6Al-4V surfaces. This was done without inducing micro-cracks or significant changes in surface chemistry within the heat affected zones. The desired 8-12 mum groove depths and widths were achieved by the control of pulse frequency, scan speed, and the lens focal length that controls spot size. The interactions between human osteosarcoma (HOS) cells and laser-grooved Ti-6Al-4V surfaces were investigated after 48 hours of cell culture. The cell behavior, including cell spreading, alignment and adhesion, was elucidated using scanning electronic microscopy (SEM), immuno-fluorescence staining and enzymatic detachment. Contact guidance was shown to increase as grooved spacing decreased. For the range of micro-groove geometries studied, micro-grooves with groove spacings of 20 mum provided the best combination of cell orientation and adhesion. Short-term adhesion experiments (15 mins to 1 day) also revealed that there is a positive correlation between cell orientation and cell adhesion. Contact guidance on the micro-grooved surfaces is shown to be enhanced by nano- and micro-scale asperities that provide sites for the attachment of lamellopodia during cell locomotion and spreading. Contact guidance is also promoted by the geometrical confinement provided by laser grooves. An experimental study of initial cell spreading and ingrowth into Ti-6Al-4V porous structures was also carried out on porous structures with different pore sizes and geometries. A combination of SEM, the tetrazolium salt (MTT) colorimetric assay and enzymatic detachment were used to study cell spreading and adhesion. The extent of cell ingrowth, pore coverage, cell adhesion and proliferation was observed to increase with decreasing pore size. It was found that fiber geometries provided guidance for cell spreading along the fiber directions. However, the larger gaps in fiber geometries made pore bridging difficult. Finally, this dissertation presents an in vivo study of the combined effects of laser microgrooving and RGD-coating on the osseointegration of implanted Ti-6Al-4V pins. Both histological and biomechanical results show that the combination of laser microgrooving and RGD-coating results in improved osseointegration over the control surfaces. All the above findings have important implications for future orthopedic and dental implant design.

Interactions of Entamoeba Histolytica with Host Cells in the Gut Mucosa,

DTIC Science & Technology

1977-01-01

amebae were observed close to the cecal epithelium, the apposing epithelial cell was found to have protruded in the lumen and making contact with the... amebae . Such cells often had become detached from their basal lamina and intercellular tight junctions, affording spaces through which amebae invaded the...mucosa. Other epithelial cells in close proximity to amebae showed degenerative changes characterized by swelling of mitochondria and endoplasmic

Retrobulbar optic neuritis and rhegmatogenous retinal detachment in a fourteen-year-old girl with retinitis pigmentosa sine pigmento.

PubMed

Hatta, M; Hayasaka, S; Kato, T; Kadoi, C

2000-01-01

A 14-year-old girl complained of a sudden decrease in right visual acuity. The patient had night blindness, a mottled retina but no pigments, extinguished scotopic electroretinographic response, central scotoma in the right eye and rhegmatogenous retinal detachment. She had initially received laser photocoagulation around the retinal tear and then corticosteroid therapy, cryoretinopexy and segmental buckling. Her right visual acuity increased to 1.0. The association of retinitis pigmentosa sine pigmento, retrobulbar optic neuritis and rhegmatogenous retinal detachment, as demonstrated in our patient, may be uncommon. Copyright 2000 S. Karger AG, Basel

Lamin A/C deficiency reduces circulating tumor cell resistance to fluid shear stress

PubMed Central

Denais, Celine; Chan, Maxine F.; Wang, Zhexiao; Lammerding, Jan

2015-01-01

Metastasis contributes to over 90% of cancer-related deaths and is initiated when cancer cells detach from the primary tumor, invade the basement membrane, and enter the circulation as circulating tumor cells (CTCs). While metastasis is viewed as an inefficient process with most CTCs dying within the bloodstream, it is evident that some CTCs are capable of resisting hemodynamic shear forces to form secondary tumors in distant tissues. We hypothesized that nuclear lamins A and C (A/C) act as key structural components within CTCs necessary to resist destruction from elevated shear forces of the bloodstream. Herein, we show that, compared with nonmalignant epithelial cells, tumor cells are resistant to elevated fluid shear forces in vitro that mimic those within the bloodstream, as evidenced by significant decreases in cellular apoptosis and necrosis. Knockdown of lamin A/C significantly reduced tumor cell resistance to fluid shear stress, with significantly increased cell death compared with parental tumor cell and nontargeting controls. Interestingly, lamin A/C knockdown increased shear stress-induced tumor cell apoptosis, but did not significantly affect cellular necrosis. These data demonstrate that lamin A/C is an important structural component that enables tumor cell resistance to fluid shear stress-mediated death in the bloodstream, and may thus facilitate survival and hematogenous metastasis of CTCs. PMID:26447202

Hormonal regulation of growth and life span of bullfrog tadpole tail epidermal cells cultured in vitro.

PubMed

Nishikawa, A; Yoshizato, K

1986-02-01

Epidermal cells were dissociated from tails of the bullfrog tadpole, Rana catesbeiana, and cultured to investigate their response to steroid and thyroid hormones. Charcoal-treated serum (CTS) was used in the growth medium when cells were to be grown in the absence of steroid and thyroid hormones. The cells could be maintained for 2 weeks with a small increase in cell number in medium that contained CTS (CTS medium). Addition of cortisol to CTS medium increased both cellular attachment to the culture dishes and the proliferation of the attached cells with an optimum concentration of 5 X 10(-7) M. The cells remained viable and attached for at least a week. Cortisol stimulated the rate of protein synthesis 1.8-fold but did not alter the rate of DNA synthesis. The cells did not proliferate in the medium containing triiodothyronine (T3) and detached themselves from the dish within 5 days, which occurred in a dose-dependent manner with a maximum effect at 10(-8) M. It drastically decreased the rate of DNA synthesis but did not influence the rate of protein synthesis. These responses of cells to cortisol and T3 may reflect growth and death of tail epidermal cells in vivo at metamorphosis.

A male contraceptive targeting germ cell adhesion.

PubMed

Mruk, Dolores D; Wong, Ching-Hang; Silvestrini, Bruno; Cheng, C Yan

2006-11-01

Throughout spermatogenesis, developing germ cells remain attached to Sertoli cells via testis-specific anchoring junctions. If adhesion between these cell types is compromised, germ cells detach from the seminiferous epithelium and infertility often results. Previously, we reported that Adjudin is capable of inducing germ cell loss from the epithelium. In a small subset of animals, however, oral administration of Adjudin (50 mg per kg body weight (b.w.) for 29 d) resulted in adverse effects such as liver inflammation and muscle atrophy. Here, we report a novel approach in which Adjudin is specifically targeted to the testis by conjugating Adjudin to a recombinant follicle-stimulating hormone (FSH) mutant, which serves as its 'carrier'. Using this approach, infertility was induced in adult rats when 0.5 microg Adjudin per kg b.w. was administered intraperitoneally, which was similar to results when 50 mg per kg b.w. was given orally. This represents a substantial increase in Adjudin's selectivity and efficacy as a male contraceptive.

Numerical simulations used for a validity check on the laser induced photo-detachment diagnostic method in electronegative plasmas

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

Oudini, N.; Taccogna, F.; Bendib, A.

2014-06-15

Laser photo-detachment is used as a method to measure or determine the negative ion density and temperature in electronegative plasmas. In essence, the method consists of producing an electropositive channel (negative ion free region) via pulsed laser photo-detachment within an electronegative plasma bulk. Electrostatic probes placed in this channel measure the change in the electron density. A second pulse might be used to track the negative ion recovery. From this, the negative ion density and temperature can be determined. We study the formation and relaxation of the electropositive channel via a two-dimensional Particle-In-Cell/Mote Carlo collision model. The simulation is mainlymore » carried out in a Hydrogen plasma with an electronegativity of α = 1, with a parametric study for α up to 20. The temporal and spatial evolution of the plasma potential and the electron densities shows the formation of a double layer (DL) confining the photo-detached electrons within the electropositive channel. This DL evolves into two fronts that move in the opposite directions inside and outside of the laser spot region. As a consequence, within the laser spot region, the background and photo-detached electron energy distribution function relaxes/thermalizes via collisionless effects such as Fermi acceleration and Landau damping. Moreover, the simulations show that collisional effects and the DL electric field strength might play a non-negligible role in the negative ion recovery within the laser spot region, leading to a two-temperature negative ion distribution. The latter result might have important effects in the determination of the negative ion density and temperature from laser photo detachment diagnostic.« less

Effectiveness of a dynein team in a tug of war helped by reduced load sensitivity of detachment: evidence from the study of bidirectional endosome transport in D. discoideum.

PubMed

Bhat, Deepak; Gopalakrishnan, Manoj

2012-08-01

Bidirectional cargo transport by molecular motors in cells is a complex phenomenon in which the cargo (usually a vesicle) alternately moves in retrograde and anterograde directions. In this case, teams of oppositely pulling motors (e.g., kinesin and dynein) bind to the cargo, simultaneously, and 'coordinate' their activity such that the motion consists of spells of positively and negatively directed segments, separated by pauses of varying duration. A set of recent experiments have analyzed the bidirectional motion of endosomes in the amoeba D. discoideum in detail. It was found that in between directional switches, a team of five to six dyneins stall a cargo against a stronger kinesin in a tug of war, which lasts for almost a second. As the mean detachment time of a kinesin under its stall load was also observed to be ∼1 s, we infer that the collective detachment time of the dynein assembly must also be similar. Here, we analyze this inference from a modeling perspective, using experimentally measured single-molecule parameters as inputs. We find that the commonly assumed exponential load-dependent detachment rate is inconsistent with observations, as it predicts that a five-dynein assembly will detach under its combined stall load in less than a hundredth of a second. A modified model where the load-dependent unbinding rate is assumed to saturate at stall-force level for super-stall loads gives results which are in agreement with experimental data. Our analysis suggests that the load-dependent detachment of a dynein in a team is qualitatively different at sub-stall and super-stall loads, a conclusion which is likely to have implications in other situations involving collective effects of many motors.

Numerical simulations used for a validity check on the laser induced photo-detachment diagnostic method in electronegative plasmas

NASA Astrophysics Data System (ADS)

Oudini, N.; Taccogna, F.; Bendib, A.; Aanesland, A.

2014-06-01

Laser photo-detachment is used as a method to measure or determine the negative ion density and temperature in electronegative plasmas. In essence, the method consists of producing an electropositive channel (negative ion free region) via pulsed laser photo-detachment within an electronegative plasma bulk. Electrostatic probes placed in this channel measure the change in the electron density. A second pulse might be used to track the negative ion recovery. From this, the negative ion density and temperature can be determined. We study the formation and relaxation of the electropositive channel via a two-dimensional Particle-In-Cell/Mote Carlo collision model. The simulation is mainly carried out in a Hydrogen plasma with an electronegativity of α = 1, with a parametric study for α up to 20. The temporal and spatial evolution of the plasma potential and the electron densities shows the formation of a double layer (DL) confining the photo-detached electrons within the electropositive channel. This DL evolves into two fronts that move in the opposite directions inside and outside of the laser spot region. As a consequence, within the laser spot region, the background and photo-detached electron energy distribution function relaxes/thermalizes via collisionless effects such as Fermi acceleration and Landau damping. Moreover, the simulations show that collisional effects and the DL electric field strength might play a non-negligible role in the negative ion recovery within the laser spot region, leading to a two-temperature negative ion distribution. The latter result might have important effects in the determination of the negative ion density and temperature from laser photo detachment diagnostic.

In vitro model to study the effects of matrix stiffening on Ca2+ handling and myofilament function in isolated adult rat cardiomyocytes

PubMed Central

Najafi, Aref; Fontoura, Dulce; Valent, Erik; Goebel, Max; Kardux, Kim; Falcão‐Pires, Inês; van der Velden, Jolanda

2017-01-01

Key points This paper describes a novel model that allows exploration of matrix‐induced cardiomyocyte adaptations independent of the passive effect of matrix rigidity on cardiomyocyte function.Detachment of adult cardiomyocytes from the matrix enables the study of matrix effects on cell shortening, Ca2+ handling and myofilament function.Cell shortening and Ca2+ handling are altered in cardiomyocytes cultured for 24 h on a stiff matrix.Matrix stiffness‐impaired cardiomyocyte contractility is reversed upon normalization of extracellular stiffness.Matrix stiffness‐induced reduction in unloaded shortening is more pronounced in cardiomyocytes isolated from obese ZSF1 rats with heart failure with preserved ejection fraction compared to lean ZSF1 rats. Abstract Extracellular matrix (ECM) stiffening is a key element of cardiac disease. Increased rigidity of the ECM passively inhibits cardiac contraction, but if and how matrix stiffening also actively alters cardiomyocyte contractility is incompletely understood. In vitro models designed to study cardiomyocyte–matrix interaction lack the possibility to separate passive inhibition by a stiff matrix from active matrix‐induced alterations of cardiomyocyte properties. Here we introduce a novel experimental model that allows exploration of cardiomyocyte functional alterations in response to matrix stiffening. Adult rat cardiomyocytes were cultured for 24 h on matrices of tuneable stiffness representing the healthy and the diseased heart and detached from their matrix before functional measurements. We demonstrate that matrix stiffening, independent of passive inhibition, reduces cell shortening and Ca2+ handling but does not alter myofilament‐generated force. Additionally, detachment of adult cultured cardiomyocytes allowed the transfer of cells from one matrix to another. This revealed that stiffness‐induced cardiomyocyte changes are reversed when matrix stiffness is normalized. These matrix stiffness‐induced changes in cardiomyocyte function could not be explained by adaptation in the microtubules. Additionally, cardiomyocytes isolated from stiff hearts of the obese ZSF1 rat model of heart failure with preserved ejection fraction show more pronounced reduction in unloaded shortening in response to matrix stiffening. Taken together, we introduce a method that allows evaluation of the influence of ECM properties on cardiomyocyte function separate from the passive inhibitory component of a stiff matrix. As such, it adds an important and physiologically relevant tool to investigate the functional consequences of cardiomyocyte–matrix interactions. PMID:28485491

Podocyte-specific RAP1GAP expression contributes to focal segmental glomerulosclerosis–associated glomerular injury

PubMed Central

Potla, Uma; Ni, Jie; Vadaparampil, Justin; Yang, Guozhe; Leventhal, Jeremy S.; Campbell, Kirk N.; Chuang, Peter Y.; Morozov, Alexei; He, John C.; D’Agati, Vivette D.; Klotman, Paul E.; Kaufman, Lewis

2014-01-01

Injury to the specialized epithelial cells of the glomerulus (podocytes) underlies the pathogenesis of all forms of proteinuric kidney disease; however, the specific genetic changes that mediate podocyte dysfunction after injury are not fully understood. Here, we performed a large-scale insertional mutagenic screen of injury-resistant podocytes isolated from mice and found that increased expression of the gene Rap1gap, encoding a RAP1 activation inhibitor, ameliorated podocyte injury resistance. Furthermore, injured podocytes in murine models of disease and kidney biopsies from glomerulosclerosis patients exhibited increased RAP1GAP, resulting in diminished glomerular RAP1 activation. In mouse models, podocyte-specific inactivation of Rap1a and Rap1b induced massive glomerulosclerosis and premature death. Podocyte-specific Rap1a and Rap1b haploinsufficiency also resulted in severe podocyte damage, including features of podocyte detachment. Over-expression of RAP1GAP in cultured podocytes induced loss of activated β1 integrin, which was similarly observed in kidney biopsies from patients. Furthermore, preventing elevation of RAP1GAP levels in injured podocytes maintained β1 integrin–mediated adhesion and prevented cellular detachment. Taken together, our findings suggest that increased podocyte expression of RAP1GAP contributes directly to podocyte dysfunction by a mechanism that involves loss of RAP1-mediated activation of β1 integrin. PMID:24642466

Anti-trypanosomal activity of cationic N-heterocyclic carbene gold(I) complexes.

PubMed

Winter, Isabel; Lockhauserbäumer, Julia; Lallinger-Kube, Gertrud; Schobert, Rainer; Ersfeld, Klaus; Biersack, Bernhard

2017-06-01

Two gold(I) N-heterocyclic carbene complexes 1a and 1b were tested for their anti-trypanosomal activity against Trypanosoma brucei parasites. Both gold compounds exhibited excellent anti-trypanosomal activity (IC 50 =0.9-3.0nM). The effects of the gold complexes 1a and 1b on the T. b. brucei cytoskeleton were evaluated. Rapid detachment of the flagellum from the cell body occurred after treatment with the gold complexes. In addition, a quick and complete degeneration of the parasitic cytoskeleton was induced by the gold complexes, only the microtubules of the detached flagellum remained intact. Both gold compounds 1a and 1b feature selective anti-trypanosomal agents and were distinctly more active against T. b. brucei cells than against human HeLa cells. Thus, the gold complexes 1a and 1b feature promising drug candidates for the treatment of trypanosome infections such as sleeping sickness (human African Trypanosomiasis caused by Trypanosoma brucei parasites). Copyright © 2017 Elsevier B.V. All rights reserved.

Network theory inspired analysis of time-resolved expression data reveals key players guiding P. patens stem cell development.

PubMed

Busch, Hauke; Boerries, Melanie; Bao, Jie; Hanke, Sebastian T; Hiss, Manuel; Tiko, Theodhor; Rensing, Stefan A

2013-01-01

Transcription factors (TFs) often trigger developmental decisions, yet, their transcripts are often only moderately regulated and thus not easily detected by conventional statistics on expression data. Here we present a method that allows to determine such genes based on trajectory analysis of time-resolved transcriptome data. As a proof of principle, we have analysed apical stem cells of filamentous moss (P. patens) protonemata that develop from leaflets upon their detachment from the plant. By our novel correlation analysis of the post detachment transcriptome kinetics we predict five out of 1,058 TFs to be involved in the signaling leading to the establishment of pluripotency. Among the predicted regulators is the basic helix loop helix TF PpRSL1, which we show to be involved in the establishment of apical stem cells in P. patens. Our methodology is expected to aid analysis of key players of developmental decisions in complex plant and animal systems.

Sediment detachment and transport processes associated with internal erosion of soil pipes: Often overlooked processes of gully erosion

USDA-ARS?s Scientific Manuscript database

Subsurface flow can be an important process in gully erosion through its impact on decreasing soil cohesion and erosion resistance as soil water content or pressure increases and more directly by the effects of seepage forces on particle detachment and piping. The development of perched water tables...

Synthesis and Characterization of Poly(Ethylene Glycol) Based Thermo-Responsive Hydrogels for Cell Sheet Engineering.

PubMed

Son, Kuk Hui; Lee, Jin Woo

2016-10-20

The swelling properties and thermal transition of hydrogels can be tailored by changing the hydrophilic-hydrophobic balance of polymer networks. Especially, poly( N -isopropylacrylamide) (PNIPAm) has received attention as thermo-responsive hydrogels for tissue engineering because its hydrophobicity and swelling property are transited around body temperature (32 °C). In this study, we investigated the potential of poly(ethylene glycol) diacrylate (PEGDA) as a hydrophilic co-monomer and crosslinker of PNIPAm to enhance biological properties of PNIPAm hydrogels. The swelling ratios, lower critical solution temperature (LCST), and internal pore structure of the synthesized p(NIPAm- co -PEGDA) hydrogels could be varied with changes in the molecular weight of PEGDA and the co-monomer ratios (NIPAm to PEGDA). We found that increasing the molecular weight of PEGDA showed an increase of pore sizes and swelling ratios of the hydrogels. In contrast, increasing the weight ratio of PEGDA under the same molecular weight condition increased the crosslinking density and decreased the swelling ratios of the hydrogels. Further, to evaluate the potential of these hydrogels as cell sheets, we seeded bovine chondrocytes on the p(NIPAm- co -PEGDA) hydrogels and observed the proliferation of the seed cells and their detachment as a cell sheet upon a decrease in temperature. Based on our results, we confirmed that p(NIPAm- co -PEGDA) hydrogels could be utilized as cell sheets with enhanced cell proliferation performance.

Annexin A2 in Proliferative Vitreoretinopathy

DTIC Science & Technology

2017-10-01

cells , leading to formation of an epiretinal membrane, retinal detachment, and loss of vision. At present, there are no reliable means of...type versus annexin A2- deficient mice, [2] define the role of A2 in the function of activated macrophages and RPE cells in PVR, and [3] examine the...expression is needed in both macrophages and RPE cells , and that A2 is extensively expressed within cells of epiretinal membranes in human PVR. Our

Extracting archaeal populations from iron oxidizing systems

NASA Astrophysics Data System (ADS)

Whitmore, L. M.; Hutchison, J.; Chrisler, W.; Jay, Z.; Moran, J.; Inskeep, W.; Kreuzer, H.

2013-12-01

Unique environments in Yellowstone National Park offer exceptional conditions for studying microorganisms in extreme and constrained systems. However, samples from some extreme systems often contain inorganic components that pose complications during microbial and molecular analysis. Several archaeal species are found in acidic, geothermal ferric-oxyhydroxide mats; these species have been shown to adhere to mineral surfaces in flocculated colonies. For optimal microbial analysis, (microscopy, flow cytometry, genomic extractions, proteomic analysis, stable isotope analysis, and others), improved techniques are needed to better facilitate cell detachment and separation from mineral surfaces. As a requirement, these techniques must preserve cell structure while simultaneously minimizing organic carryover to downstream analysis. Several methods have been developed for removing sediments from mixed prokaryotic populations, including ultra-centrifugation, nycodenz gradient, sucrose cushions, and cell straining. In this study we conduct a comparative analysis of mechanisms used to detach archaeal cell populations from the mineral interface. Specifically, we evaluated mechanical and chemical approaches for cell separation and homogenization. Methods were compared using confocal microscopy, flow cytometry analyses, and real-time PCR detection. The methodology and approaches identified will be used to optimize biomass collection from environmental specimens or isolates grown with solid phases.

Suspension state increases reattachment of breast cancer cells by up-regulating lamin A/C.

PubMed

Zhang, Xiaomei; Lv, Yonggang

2017-12-01

Extravasation is a rate-limiting step of tumor metastasis, for which adhesion to endothelium of circulating tumor cells (CTCs) is the prerequisite. The suspension state of CTCs undergoing detachment from primary tumor is a persistent biomechanical cue, which potentially regulates the biophysical characteristics and cellular behaviors of tumor cells. In this study, breast tumor cells MDA-MB-231 in suspension culture condition were used to investigate the effect of suspension state on reattachment of CTCs. Our study demonstrated that suspension state significantly increased the adhesion ability of breast tumor cells. In addition, suspension state markedly promoted the formation of stress fibers and focal adhesions and reduced the motility in reattached breast cancer cells. Moreover, lamin A/C was reversibly accumulated at posttranscriptional level under suspension state, improving the cell stiffness of reattached breast cancer cells. Disruption of actin cytoskeleton by cytochalasin D caused lamin A/C accumulation. Conversely, decreasing actomyosin contraction by ROCK inhibitor Y27632 reduced lamin A/C level. Knocking down lamin A/C weakened the suspension-induced increase of adhesion, and also abolished the suspension-induced decrease of motility and increase of stress fibers and focal adhesion in reattaching tumor cells, suggesting a crucial role of lamin A/C. In conclusion, it was demonstrated that suspension state promoted the reattachment of breast tumor cells by up-regulating lamin A/C via cytoskeleton disruption. These findings highlight the important role of suspension state for tumor cells in tumor metastasis. Copyright © 2017 Elsevier B.V. All rights reserved.

Dihydroartemisinin induces endothelial cell anoikis through the activation of the JNK signaling pathway

PubMed Central

Zhang, Jiao; Guo, Ling; Zhou, Xia; Dong, Fengyun; Li, Liqun; Cheng, Zuowang; Xu, Yinghua; Liang, Jiyong; Xie, Qi; Liu, Ju

2016-01-01

Angiogenesis is required for the growth and metastasis of solid tumors. The anti-malarial agent dihydroartemisinin (DHA) demonstrates potent anti-angiogenic activity, but the underlying molecular mechanisms are not yet fully understood. During the process of angiogenesis, endothelial cells migrating from existing capillaries may undergo programmed cell death after detaching from the extracellular matrix, a process that is defined as anchorage-dependent apoptosis or anoikis. In the present study, DHA-induced cell death was compared in human umbilical vein endothelial cells (HUVECs) cultured in suspension and attached to culture plates. In suspended HUVECs, the cell viability was decreased and apoptosis was increased with the treatment of 50 µM DHA for 5 h, while the same treatment did not affect the attached HUVECs. In addition, 50 µM DHA increased the phosphorylation of c-Jun N-terminal kinase (JNK) in suspended HUVECs, but not in attached HUVECs, for up to 5 h of treatment. The JNK inhibitor, SP600125, reversed DHA-induced cell death in suspended HUVECs, suggesting that the JNK pathway may mediate DHA-induced endothelial cell anoikis. The data from the present study indicates a novel mechanism for understanding the anti-angiogenic effects of DHA, which may be used as a component for chemotherapy. PMID:27602117

Measurement of He neutral temperature in detached plasmas using laser absorption spectroscopy

NASA Astrophysics Data System (ADS)

Aramaki, M.; Tsujihara, T.; Kajita, S.; Tanaka, H.; Ohno, N.

2018-01-01

The reduction of the heat load onto plasma-facing components by plasma detachment is an inevitable scheme in future nuclear fusion reactors. Since the control of the plasma and neutral temperatures is a key issue to the detached plasma generation, we have developed a laser absorption spectroscopy system for the metastable helium temperature measurements and used together with a previously developed laser Thomson scattering system for the electron temperature and density measurements. The thermal relaxation process between the neutral and the electron in the detached plasma generated in the linear plasma device, NAGDIS-II was studied. It is shown that the electron temperature gets close to the neutral temperature by increasing the electron density. On the other hand, the pressure dependence of electron and neutral temperatures shows the cooling effect by the neutrals. The possibility of the plasma fluctuation measurement using the fluctuation in the absorption signal is also shown.

Histopathological and immunohistochemical findings associated with a null mutation in the Norrie disease gene.

PubMed

Schroeder, B; Hesse, L; Brück, W; Gal, A

1997-06-01

To determine the clinical, histopathological, and immunohistochemical ocular changes associated with a null mutation in the Norrie disease protein (NDP) gene. Tissue from a six-month-old boy with bilateral retrolental membranes and retinal detachment was obtained during vitreoretinal surgery. Histological sections were stained immunohistochemically with specific antibodies. No eye diseases with severe visual impairment or blindness were reported in the parents and their families. The NDP gene was analyzed by standard molecular genetic methods. A severe reduction in the number of retinal ganglion cells and a largely disarranged and hypoplastic inner nuclear layer were visible in the tissue specimen. Areas of the tissue with advanced pathology displayed massive fibrovascular proliferation in the vitreous cavity. Shrinkage and traction resulted in folding and detachment of the outer retina. Immunohistochemical reactivity for MIB(1) antigen demonstrated many proliferating cells in the vitreous, but no proliferative activity in the neuroretina. Retinal neurons showed a high grade of differentiation and expressed uniformly neuron-specific enolase and synaptophysin. A 1-base pair insertion (544/545insA) in the NDP gene was found in the affected boy. This mutation predicts a 'functional null-allele' due to a shift in the reading frame and, thus, a premature termination of mRNA translation after 55 instead of 133 amino acids. Loss of function of the NDP gene causes marked hypoplasia of the inner retinal cell layers and fibrovascular proliferation in the vitreous cavity, leading to retinal folding and detachment. The NDP therefore seems to play a critical role in terminal differentiation of the inner retinal cell layers and establishment and maintaining of anti-proliferative cellular interactions in the vitreous.

Induction of morphological changes in death-induced cancer cells monitored by holographic microscopy.

PubMed

El-Schich, Zahra; Mölder, Anna; Tassidis, Helena; Härkönen, Pirkko; Falck Miniotis, Maria; Gjörloff Wingren, Anette

2015-03-01

We are using the label-free technique of holographic microscopy to analyze cellular parameters including cell number, confluence, cellular volume and area directly in the cell culture environment. We show that death-induced cells can be distinguished from untreated counterparts by the use of holographic microscopy, and we demonstrate its capability for cell death assessment. Morphological analysis of two representative cell lines (L929 and DU145) was performed in the culture flasks without any prior cell detachment. The two cell lines were treated with the anti-tumour agent etoposide for 1-3days. Measurements by holographic microscopy showed significant differences in average cell number, confluence, volume and area when comparing etoposide-treated with untreated cells. The cell volume of the treated cell lines was initially increased at early time-points. By time, cells decreased in volume, especially when treated with high doses of etoposide. In conclusion, we have shown that holographic microscopy allows label-free and completely non-invasive morphological measurements of cell growth, viability and death. Future applications could include real-time monitoring of these holographic microscopy parameters in cells in response to clinically relevant compounds. Copyright © 2015 Elsevier Inc. All rights reserved.

A general description of detachment for multidimensional modelling of biofilms.

PubMed

Xavier, Joao de Bivar; Picioreanu, Cristian; van Loosdrecht, Mark C M

2005-09-20

A general method for describing biomass detachment in multidimensional biofilm modelling is introduced. Biomass losses from processes acting on the entire surface of the biofilm, such as erosion, are modelled using a continuous detachment speed function F(det). Discrete detachment events, i.e. sloughing, are implicitly derived from simulations. The method is flexible to allow F(det) to take several forms, including expressions dependent on any state variables such as the local biofilm density. This methodology for biomass detachment was integrated with multidimensional (2D and 3D) particle-based multispecies biofilm models by using a novel application of the level set method. Application of the method is illustrated by trends in the dynamics of biofilms structure and activity derived from simulations performed on a simple model considering uniform biomass (case study I) and a model discriminating biomass composition in heterotrophic active mass, extracellular polymeric substances (EPS) and inert mass (case study II). Results from case study I demonstrate the effect of applied detachment forces as a fundamental factor influencing steady-state biofilm activity and structure. Trends from experimental observations reported in literature were correctly described. For example, simulation results indicated that biomass sloughing is reduced when erosion forces are increased. Case study II illustrates the application of the detachment methodology to systems with non-uniform biomass composition. Simulations carried out at different bulk concentrations of substrate show changes in biofilm structure (in terms of shape, density and spatial distribution of biomass components) and activity (in terms of oxygen and substrate consumption) as a consequence of either oxygen-limited or substrate-limited growth. (c) 2005 Wiley Periodicals, Inc.

PEG-detachable lipid-polymer hybrid nanoparticle for delivery of chemotherapy drugs to cancer cells.

PubMed

Du, Jiang-bo; Song, Yan-feng; Ye, Wei-liang; Cheng, Ying; Cui, Han; Liu, Dao-zhou; Liu, Miao; Zhang, Bang-le; Zhou, Si-yuan

2014-08-01

The experiment aimed to increase the drug-delivery efficiency of poly-lactic-co-glycolic acid (PLGA) nanoparticles. Lipid-polymer hybrid nanoparticles (LPNs-1) were prepared using PLGA as a hydrophobic core and FA-PEG-hyd-DSPE as an amphiphilic shell. Uniform and spherical nanoparticles with an average size of 185 nm were obtained using the emulsification solvent evaporation method. The results indicated that LPNs-1 showed higher drug loading compared with naked PLGA nanoparticles (NNPs). Drug release from LPNs-1 was faster in an acidic environment than in a neutral environment. LPNs-1 showed higher cytotoxicity on KB cells, A549 cells, MDA-MB-231 cells, and MDA-MB-231/ADR cells compared with free doxorubicin (DOX) and NNPs. The results also showed that, compared with free DOX and NNPs, LPNs-1 delivered more DOX to the nuclear of KB cells and MDA-MB-231/ADR cells. LPNs-1 induced apoptosis in KB cells and MDA-MB-231/ADR cells in a dose-dependent manner. The above data indicated that DOX-loaded LPNs-1 could kill not only normal tumor cells but also drug-resistant tumor cells. These results indicated that modification of PLGA nanoparticles with FA-PEG-hyd-DSPE could considerably increase the drug-delivery efficiency and LPNs-1 had potential in the delivery of chemotherapeutic agents in the treatment of cancer.

The ``Swiss cheese'' instability of bacterial biofilms

NASA Astrophysics Data System (ADS)

Jang, Hongchul; Rusconi, Roberto; Stocker, Roman

2012-11-01

Bacteria often adhere to surfaces, where they develop polymer-encased communities (biofilms) that display dramatic resistance to antibiotic treatment. A better understanding of cell detachment from biofilms may lead to novel strategies for biofilm disruption. Here we describe a new detachment mode, whereby a biofilm develops a nearly regular array of ~50-100 μm holes. Using surface-treated microfluidic devices, we create biofilms of controlled shape and size. After the passage of an air plug, the break-up of the residual thin liquid film scrapes and rearranges bacteria on the surface, such that a ``Swiss cheese'' pattern is left in the residual biofilm. Fluorescent staining of the polymeric matrix (EPS) reveals that resistance to cell dislodgement correlates with local biofilm age, early settlers having had more time to hunker down. Because few survivors suffice to regrow a biofilm, these results point at the importance of considering microscale heterogeneity in assessing the effectiveness of biofilm removal strategies.

Rapid fabrication of detachable three-dimensional tissues by layering of cell sheets with heating centrifuge.

PubMed

Haraguchi, Yuji; Kagawa, Yuki; Hasegawa, Akiyuki; Kubo, Hirotsugu; Shimizu, Tatsuya

2018-01-18

Confluent cultured cells on a temperature-responsive culture dish can be harvested as an intact cell sheet by decreasing temperature below 32°C. A three-dimensional (3-D) tissue can be fabricated by the layering of cell sheets. A resulting 3-D multilayered cell sheet-tissue on a temperature-responsive culture dish can be also harvested without any damage by only temperature decreasing. For shortening the fabrication time of the 3-D multilayered constructs, we attempted to layer cell sheets on a temperature-responsive culture dish with centrifugation. However, when a cell sheet was attached to the culture surface with a conventional centrifuge at 22-23°C, the cell sheet hardly adhere to the surface due to its noncell adhesiveness. Therefore, in this study, we have developed a heating centrifuge. In centrifugation (55g) at 36-37°C, the cell sheet adhered tightly within 5 min to the dish without significant cell damage. Additionally, centrifugation accelerated the cell sheet-layering process. The heating centrifugation shortened the fabrication time by one-fifth compared to a multilayer tissue fabrication without centrifugation. Furthermore, the multilayered constructs were finally detached from the dishes by decreasing temperature. This rapid tissue-fabrication method will be used as a valuable tool in the field of tissue engineering and regenerative therapy. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018. © 2018 American Institute of Chemical Engineers.

Mechanics of membrane-cytoskeleton attachment in Paramecium

NASA Astrophysics Data System (ADS)

Campillo, C.; Jerber, J.; Fisch, C.; Simoes-Betbeder, M.; Dupuis-Williams, P.; Nassoy, P.; Sykes, C.

2012-12-01

In this paper we assess the role of the protein MKS1 (Meckel syndrome type 1) in the cortical membrane mechanics of the ciliated protist Paramecium. This protein is known to be crucial in the process of cilium formation, and we investigate its putative role in membrane-cytoskeleton attachment. Therefore, we compare cells where the gene coding for MKS1 is silenced to wild-type cells. We found that scanning electron microscopy observation of the cell surface reveals a cup-like structure in wild-type cells that is lost in silenced cells. Since this structure is based on the underlying cytoskeleton, one hypothesis to explain this observation is a disruption of membrane attachment to the cytoskeleton in the absence of MKS1 that should affect plasma membrane mechanics. We test this by probing the mechanics of wild-type and silenced cells by micropipette aspiration. Strikingly, we observe that, at the same aspiration pressure, the membrane of silenced cells is easily aspirated by the micropipette whereas that of wild-type cells enters only at a moderate velocity, an effect that suggests a detachment of the membrane from the underlying cytoskeleton in silenced cells. We quantify this detachment by measuring the deformation of the cell cortex and the rate of cell membrane entry in the micropipette. This study offers a new perspective for the characterization of membrane-cytoskeleton attachment in protists and paves the way for a better understanding of the role of membrane-cortex attachment in cilium formation.

Manufacturing human mesenchymal stem cells at clinical scale: process and regulatory challenges.

PubMed

Jossen, Valentin; van den Bos, Christian; Eibl, Regine; Eibl, Dieter

2018-05-01

Human mesenchymal stem cell (hMSC)-based therapies are of increasing interest in the field of regenerative medicine. As economic considerations have shown, allogeneic therapy seems to be the most cost-effective method. Standardized procedures based on instrumented single-use bioreactors have been shown to provide billion of cells with consistent product quality and to be superior to traditional expansions in planar cultivation systems. Furthermore, under consideration of the complex nature and requirements of allogeneic hMSC-therapeutics, a new equipment for downstream processing (DSP) was successfully evaluated. This mini-review summarizes both the current state of the hMSC production process and the challenges which have to be taken into account when efficiently producing hMSCs for the clinical scale. Special emphasis is placed on the upstream processing (USP) and DSP operations which cover expansion, harvesting, detachment, separation, washing and concentration steps, and the regulatory demands.

Effect of Dissolution Kinetics on Feature Size in Dip-Pen Nanolithography

NASA Astrophysics Data System (ADS)

Weeks, B. L.; Noy, A.; Miller, A. E.; de Yoreo, J. J.

2002-06-01

We have investigated the effects of humidity, tip speed, and dwell time on feature size during dip pen nanolithography. Our results indicate a transition between two distinct deposition regimes occurs at a dwell time independent of humidity. While feature size increases with humidity, the relative increase is independent of dwell time. The results are described by a model that accounts for detachment and reattachment at the tip. The model suggests that, at short dwell times (high speed), the most important parameter controlling the feature size is the activation energy for thiol detachment.

Overcoming Multidrug Resistance through the GLUT1-Mediated and Enzyme-Triggered Mitochondrial Targeting Conjugate with Redox-Sensitive Paclitaxel Release.

PubMed

Ma, Pengkai; Chen, Jianhua; Bi, Xinning; Li, Zhihui; Gao, Xing; Li, Hongpin; Zhu, Hongyu; Huang, Yunfang; Qi, Jing; Zhang, Yujie

2018-04-18

Multidrug resistance (MDR) is thought to be the major obstacle leading to the failure of paclitaxel (PTX) chemotherapy. To solve this problem, a glucose transporter-mediated and matrix metalloproteinase 2 (MMP2)-triggered mitochondrion-targeting conjugate [glucose-polyethylene glycol (PEG)-peptide-triphenylphosponium-polyamidoamine (PAMAM)-PTX] composed of a PAMAM dendrimer and enzymatic detachable glucose-PEG was constructed for mitochondrial delivery of PTX. The conjugate was characterized by a 30 nm sphere particle, MMP2-sensitive PEG outer layer detachment from PAMAM, and glutathione (GSH)-sensitive PTX release. It showed higher cellular uptake both in glucose transporter 1 (GLUT1) overexpressing MCF-7/MDR monolayer cell (2D) and multicellular tumor spheroids (3D). The subcellular location study showed that it could specifically accumulate in the mitochondria. Moreover, it exhibited higher cytotoxicity against MCF-7/MDR cells, which significantly reverse the MDR of MCF-7/MDR cells. The MDR reverse might be caused by reducing the ATP content through destroying the mitochondrial membrane as well as by down-regulating P-gp expression. In vivo imaging and tissue distribution indicated more conjugate accumulated in the tumor of the tumor-bearing mice model. Consequently, the conjugate showed better tumor inhibition rate and lower body weight loss, which demonstrated that it possessed high efficiency and low toxicity. This study provides glucose-mediated GLUT targeting, MMP2-responsive PEG detachment, triphenylphosponium-mediated mitochondria targeting, and a GSH-sensitive intracellular drug release conjugate that has the potential to be exploited for overcoming MDR of PTX.

Documentation of normal stratum corneum scaling in an average population: features of differences among age, ethnicity and body site.

PubMed

Chu, M; Kollias, N

2011-03-01

Scaling skin involves an imbalance between cell proliferation and desquamation, resulting in partially detached corneocytes at the stratum corneum (SC) surface that become visible as they scatter light. The purpose of this study was to document scaling skin with no associated pathology, to estimate the range of normal corneocyte detachment in the average population, and to determine if age, pigmentation and/or body sites of different exposures contribute to differences observed in the SC. Healthy African-American and Caucasian female subjects (n = 151) from a typical central New Jersey population, aged between 14 and 75 years, were evaluated on the dorsal forearm and upper inner arm. Dermatoscopy and adhesive tape were used to evaluate the appearance and adhesion of surface corneocytes. Transepidermal water loss and conductivity were measured to assess water-handling properties of the SC. Measurements were conducted during the winter. Corneocyte detachment observed with dermatoscopy became more prevalent with age and was more severe on the dorsal forearm and in Caucasian subjects. The distribution of the amount of corneocyte removal with adhesive tape increased with age. The range of values was larger in the dorsal forearm than the upper inner arm and was greater in Caucasian subjects than African-American subjects. Minimal changes were observed for water-handling properties. The architecture of the outer SC appears different between ages, body sites of different exposures, and individuals of different pigmentation groups, but minimal differences in water-handling properties are observed. © 2011 The Authors. BJD © 2011 British Association of Dermatologists.

Efficacy of Arnica Echinacea powder in umbilical cord care in a large cohort study.

PubMed

Perrone, Serafina; Coppi, Silvia; Coviello, Caterina; Cecchi, Sara; Becucci, Elisa; Tataranno, Maria Luisa; Buonocore, Giuseppe

2012-07-01

Today healthy newborns are discharged after 48 h-72 h of life until umbilical cord (UC) detachment. Complications due to an inappropriate management are: erythema, edema, bleeding, omphalitis and sepsis. Hence the importance of a safe, effective, easy to do, and cheap method. This study tests the effects and the efficacy of arnica echinacea powder by evaluating the time of cord detachment and the risk of side effects in a large cohort of newborns. 6323 babies were treated with Arnica Echinacea powder twice a day until cord stump detachment. Medications started in hospital ward and continued at home until stump detachment. The UC stump detachment occurred in 89.09% of newborns during the first 4 days of life. This percentage increase to 96.13% at 6 days. Our study demonstrates the efficacy and the safety of arnica echinacea in UC separation. No infections or even bacterial colonizations were found. The use of arnica echinacea reduces hospital costs as a consequence of complications. In addition arnica use is well received by medical staff and parents. In conclusion due to its potential benefits, low cost and feasibility, we recommend the use of arnica echinacea powder as routine procedure in all nurseries.

Altered sensitivity to ellagic acid in neuroblastoma cells undergoing differentiation with 12-O-tetradecanoylphorbol-13-acetate and all-trans retinoic acid.

PubMed

Alfredsson, Christina Fjæraa; Rendel, Filip; Liang, Qui-Li; Sundström, Birgitta E; Nånberg, Eewa

2015-12-01

Ellagic acid has previously been reported to induce reduced proliferation and activation of apoptosis in several tumor cell lines including our own previous data from non-differentiated human neuroblastoma SH-SY5Y cells. The aim of this study was now to investigate if in vitro differentiation with the phorbol ester 12-O- tetradecanoylphorbol-13-acetate or the vitamin A derivative all-trans retinoic acid altered the sensitivity to ellagic acid in SH-SY5Y cells. The methods used were cell counting and LDH-assay for evaluation of cell number and cell death, flow cytometric analysis of SubG1- and TUNEL-analysis for apoptosis and western blot for expression of apoptosis-associated proteins. In vitro differentiation was shown to reduce the sensitivity to ellagic acid with respect to cell detachment, loss of viability and activation of apoptosis. The protective effect was phenotype-specific and most prominent in all-trans retinoic acid-differentiated cultures. Differentiation-dependent up-regulation of Bcl-2 and integrin expression is introduced as possible protective mechanisms. The presented data also point to a positive correlation between proliferative activity and sensitivity to ellagic-acid-induced cell detachment. In conclusion, the presented data emphasize the need to consider degree of neuronal differentiation and phenotype of neuroblastoma cells when discussing a potential pharmaceutical application of ellagic acid in tumor treatment. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

Modeling of detachment experiments at DIII-D

DOE PAGES

Canik, John M.; Briesemeister, Alexis R.; Lasnier, C. J.; ...

2014-11-26

Edge fluid–plasma/kinetic–neutral modeling of well-diagnosed DIII-D experiments is performed in order to document in detail how well certain aspects of experimental measurements are reproduced within the model as the transition to detachment is approached. Results indicate, that at high densities near detachment onset, the poloidal temperature profile produced in the simulations agrees well with that measured in experiment. However, matching the heat flux in the model requires a significant increase in the radiated power compared to what is predicted using standard chemical sputtering rates. Lastly, these results suggest that the model is adequate to predict the divertor temperature, provided thatmore » the discrepancy in radiated power level can be resolved.« less

Clustering T cell GM1 Lipid Rafts Increases Cellular Resistance to Shear on Fibronectin through Changes in Integrin Affinity and Cytoskeletal Dynamics

PubMed Central

Mitchell, Jason S.; Brown, Wells S.; Woodside, Darren G.; Vanderslice, Peter; McIntyre, Bradley W.

2008-01-01

Lipid rafts are small laterally mobile microdomains that are highly enriched in lymphocyte signaling molecules. GM1 gangliosides are a common lipid raft component and have been shown to be important in many T cell functions. The aggregation of specific GM1 lipid rafts can control many T cell activation events, including their novel association with T cell integrins. We found that clustering GM1 lipid rafts can regulate β1 integrin function. This was apparent through increased resistance to shear flow dependent detachment of T cells adherent to the α4β1 and α5β1 integrin ligand fibronectin (FN). Adhesion strengthening as a result of clustering GM1 enriched lipid rafts correlated with increased cellular rigidity and morphology through the localization of cortical F-actin, the resistance to shear induced cell stretching, and an increase in the surface area and symmetry of the contact area between the cell surface and adhesive substrate. Furthermore, clustering GM1 lipid rafts could initiate integrin “inside-out” signaling mechanisms. This was seen through increased integrin-cytoskeleton associations and enhanced soluble binding of FN and VCAM-1 suggesting the induction of high affinity integrin conformations. The activation of these adhesion strengthening characteristics appear to be specific for the aggregation of GM1 lipid rafts as the aggregation of the heterogeneous raft associated molecule CD59 failed to activate these functions. These findings indicate a novel mechanism to signal to β1 integrins and to activate adhesion strengthening processes. PMID:19139760

The in vitro toxicity of peritoneal dialysis fluid.

PubMed

Manuprasert, Wasin; Kanchanabuch, Sirigul; Eiam-Ong, Somchai; Kanjanabuch, Talerngsak

2011-09-01

To investigate the toxicity of peritoneal dialysis fluid (PDF) components on peritoneal changes in primary human mesothelial cell. To investigate the mechanism of changes, primary human peritoneal mesothelial cells (HPMCs) were isolated from human omental tissue and were exposed for 15 hours with the various concentrations of conventional PDF and various PDF components. The mesothelial injury was determined by calculating a ratio of supernatant and total intracellular LDH while mesothelial apoptosis was assessed and counted by positive TUNEL staining and flow cytometry, respectively. PDF caused mesothelial detachment, de-differentiation, cell injuries, and apoptosis and this depended on the concentrations of PDF. The acidic condition and high glucose concentration likely played a major role in the HPMC injuries and detachment while individual PDF component could not yield mesothelial apoptosis as severe as the whole PDF effects. Thus, the additive effects of PDF composition, instead of the effect of each component, contributed to dialysis-related HPMC damages. PDF showed concentration dependent fashion-induced HPMC injury, dedifferentiation, and apoptosis. All of the abnormalities occurred by the additive effects of PDF components.

Inter-dependent apical microtubule and actin dynamics orchestrate centrosome retention and neuronal delamination

PubMed Central

Kasioulis, Ioannis

2017-01-01

Detachment of newborn neurons from the neuroepithelium is required for correct neuronal architecture and functional circuitry. This process, also known as delamination, involves adherens-junction disassembly and acto-myosin-mediated abscission, during which the centrosome is retained while apical/ciliary membranes are shed. Cell-biological mechanisms mediating delamination are, however, poorly understood. Using live-tissue and super-resolution imaging, we uncover a centrosome-nucleated wheel-like microtubule configuration, aligned with the apical actin cable and adherens-junctions within chick and mouse neuroepithelial cells. These microtubules maintain adherens-junctions while actin maintains microtubules, adherens-junctions and apical end-foot dimensions. During neuronal delamination, acto-myosin constriction generates a tunnel-like actin-microtubule configuration through which the centrosome translocates. This movement requires inter-dependent actin and microtubule activity, and we identify drebrin as a potential coordinator of these cytoskeletal dynamics. Furthermore, centrosome compromise revealed that this organelle is required for delamination. These findings identify new cytoskeletal configurations and regulatory relationships that orchestrate neuronal delamination and may inform mechanisms underlying pathological epithelial cell detachment. PMID:29058679

AKT1 provides an essential survival signal required for differentiation and stratification of primary human keratinocytes.

PubMed

Thrash, Barry R; Menges, Craig W; Pierce, Robert H; McCance, Dennis J

2006-04-28

Keratinocyte differentiation and stratification are complex processes involving multiple signaling pathways, which convert a basal proliferative cell into an inviable rigid squame. Loss of attachment to the basement membrane triggers keratinocyte differentiation, while in other epithelial cells, detachment from the extracellular matrix leads to rapid programmed cell death or anoikis. The potential role of AKT in providing a survival signal necessary for stratification and differentiation of primary human keratinocytes was investigated. AKT activity increased during keratinocyte differentiation and was attributed to the specific activation of AKT1 and AKT2. Targeted reduction of AKT1 expression, but not AKT2, by RNA interference resulted in an abnormal epidermis in organotypic skin cultures with a thin parabasal region and a pronounced but disorganized cornified layer. This abnormal stratification was due to significant cell death in the suprabasal layers and was alleviated by caspase inhibition. Normal expression patterns of both early and late markers of keratinocyte differentiation were also disrupted, producing a poorly developed stratum corneum.

Methionine Deprivation Induces a Targetable Vulnerability in Triple-negative Breast Cancer Cells by Enhancing TRAIL Receptor-2 Expression

PubMed Central

Strekalova, Elena; Malin, Dmitry; Good, David M.; Cryns, Vincent L.

2015-01-01

Purpose Many neoplasms are vulnerable to methionine deficiency by mechanisms that are poorly understood. Because gene profiling studies have revealed that methionine depletion increases TNF-related apoptosis-inducing ligand receptor-2 (TRAIL-R2) mRNA, we postulated that methionine stress sensitizes breast cancer cells to proapoptotic TRAIL-R2 agonists. Experimental Design Human triple (ER/PR/HER2)-negative breast carcinoma cell lines were cultured in control or methionine-free media. The effects of methionine depletion on TRAIL receptor expression and sensitivity to chemotherapy or a humanized agonistic TRAIL-R2 monoclonal antibody (lexatumumab) were determined. The melanoma-associated antigen MAGED2 was silenced to delineate its functional role in sensitizing TNBC cells to methionine stress. An orthotopic TNBC model was utilized to evaluate the effects of dietary methionine deficiency, lexatumumab or the combination. Results Methionine depletion sensitized TNBC cells to lexatumumab-induced caspase activation and apoptosis by increasing TRAIL-R2 mRNA and cell surface expression. MCF-10A cells transformed by oncogenic H-Ras, but not untransformed cells, and matrix-detached TNBC cells were highly sensitive to the combination of lexatumumab and methionine depletion. Proteomics analyses revealed that MAGED2, which has been reported to reduce TRAIL-R2 expression, was suppressed by methionine stress. Silencing MAGED2 recapitulated features of methionine deprivation, including enhanced mRNA and cell surface expression of TRAIL receptors and increased sensitivity to TRAIL receptor agonists. Dietary methionine deprivation enhanced the antitumor effects of lexatumumab in an orthotopic metastatic TNBC model. Conclusion Methionine depletion exposes a targetable defect in TNBC cells by increasing TRAIL-R2 expression. Our findings provide the foundation for a clinical trial combining dietary methionine restriction and TRAIL-R2 agonists. PMID:25724522

Electron pressure balance in the SOL through the transition to detachment

DOE PAGES

McLean, A. G.; Leonard, A. W.; Makowski, M. A.; ...

2015-02-07

Upgrades to core and divertor Thomson scattering (DTS) diagnostics at DIII-D have provided measurements of electron pressure profiles in the lower divertor from attached- to fully-detached divertor plasma conditions. Detailed, multistep sequences of discharges with increasing line-averaged density were run at several levels of P inj. Strike point sweeping allowed 2D divertor characterization using DTS optimized to measure T e down to 0.5 eV. The ionization front at the onset of detachment is found to move upwards in a controlled manner consistent with the indication that scrape-off layer parallel power flux is converted from conducted to convective heat transport. Measurementsmore » of n e, T e and p e in the divertor versus Lparallel demonstrate a rapid transition from Te ≥ 15 eV to ≤3 eV occurring both at the outer strike point and upstream of the X-point. Furthermore, these observations provide a strong benchmark for ongoing modeling of divertor detachment for existing and future tokamak devices.« less

Electron pressure balance in the SOL through the transition to detachment

NASA Astrophysics Data System (ADS)

McLean, A. G.; Leonard, A. W.; Makowski, M. A.; Groth, M.; Allen, S. L.; Boedo, J. A.; Bray, B. D.; Briesemeister, A. R.; Carlstrom, T. N.; Eldon, D.; Fenstermacher, M. E.; Hill, D. N.; Lasnier, C. J.; Liu, C.; Osborne, T. H.; Petrie, T. W.; Soukhanovskii, V. A.; Stangeby, P. C.; Tsui, C.; Unterberg, E. A.; Watkins, J. G.

2015-08-01

Upgrades to core and divertor Thomson scattering (DTS) diagnostics at DIII-D have provided measurements of electron pressure profiles in the lower divertor from attached- to fully-detached divertor plasma conditions. Detailed, multistep sequences of discharges with increasing line-averaged density were run at several levels of Pinj. Strike point sweeping allowed 2D divertor characterization using DTS optimized to measure Te down to 0.5 eV. The ionization front at the onset of detachment is found to move upwards in a controlled manner consistent with the indication that scrape-off layer parallel power flux is converted from conducted to convective heat transport. Measurements of ne, Te and pe in the divertor versus Lparallel demonstrate a rapid transition from Te ⩾ 15 eV to ⩽3 eV occurring both at the outer strike point and upstream of the X-point. These observations provide a strong benchmark for ongoing modeling of divertor detachment for existing and future tokamak devices.

Long-term study of patients with congenital pit of the optic nerve and persistent macular detachment.

PubMed

Theodossiadis, G P; Panopoulos, M; Kollia, A K; Georgopoulos, G

1992-08-01

During the period 1970-87 we evaluated the changes of the optic disc, peripapillary area, detached macula and visual acuity in 16 cases with congenital pit of the optic nerve and macular detachment. The study revealed in 9 of the 16 cases (56%) an increase of the dimension of the pit or changes in its color, findings which were directly related to the duration of the macular detachment. Chorioretinal scarring, pigment migration, or both, were also noted mainly at the temporal margin of optic disc. In 5/16 cases we found during the follow-up an extension of macular elevation. In altogether 10 out of 16 cases the retinal elevation covered the larger portion of the mid-periphery temporally. In 7/16 cases the final visual acuity remained unchanged, in 9/16 cases deteriorated. The difference, however, in the latter 9 cases between initial and final visual acuity was negligible. During the follow-up period deterioration of the visual fields was also noted.

Study of the dielectric breakdown properties of CO2-O2 mixtures by considering electron detachments from negative ions

NASA Astrophysics Data System (ADS)

Zhao, Hu; Tian, Zengyao; Deng, Yunkun; Li, Xingwen; Lin, Hui

2017-12-01

The dielectric breakdown properties of CO2-O2 mixtures at different O2 concentrations and gas pressures were studied in this paper, with electron detachments from negative ions taken into consideration. The influences of the electron detachment on the reduced effective ionization coefficients αeff/N, the critical reduced electric fields (E/N)cr, the critical electron temperature Tcr, the breakdown reduced electric fields (E/N)breakdown, and the breakdown electron temperature Tbreakdown were analyzed for the CO2-O2 mixture. Based on the results, it was found that an enhancement in αeff/N and a decrease in (E/N)cr and Tcr were caused by the electron detachment, which appeared to be more significant at relatively low E/N and low gas pressures. With the increase in the pd product, both (E/N)breakdown and Tbreakdown in the CO2-O2 mixture decreased first and then tended to be a constant at relatively high pd products.

Infiltration of meteoric fluids in an extensional detachment shear zone (Kettle dome, WA, USA): How quartz dynamic recrystallization relates to fluid-rock interaction

NASA Astrophysics Data System (ADS)

Quilichini, Antoine; Siebenaller, Luc; Nachlas, William O.; Teyssier, Christian; Vennemann, Torsten W.; Heizler, Matthew T.; Mulch, Andreas

2015-02-01

We document the interplay between meteoric fluid flow and deformation processes in quartzite-dominated lithologies within a ductile shear zone in the footwall of a Cordilleran extensional fault (Kettle detachment system, Washington, USA). Across 150 m of shear zone section, hydrogen isotope ratios (δD) from synkinematic muscovite fish are constant (δD ˜ -130‰) and consistent with a meteoric fluid source. Quartz-muscovite oxygen isotope thermometry indicates equilibrium fractionation temperatures of ˜365 ± 30 °C in the lower part of the section, where grain-scale quartz deformation was dominated by grain boundary migration recrystallization. In the upper part of the section, muscovite shows increasing intragrain compositional zoning, and quartz microstructures reflect bulging recrystallization, solution-precipitation, and microcracking that developed during progressive cooling and exhumation. The preserved microstructural characteristics and hydrogen isotope fingerprints of meteoric fluids developed over a short time interval as indicated by consistent mica 40Ar/39Ar ages ranging between 51 and 50 Ma over the entire section. Pervasive fluid flow became increasingly channelized during detachment activity, leading to microstructural heterogeneity and large shifts in quartz δ18O values on a meter scale. Ductile deformation ended when brittle motion on the detachment fault rapidly exhumed the mylonitic footwall.

Bubble Generation in a Continuous Liquid Flow Under Reduced Gravity Conditions

NASA Technical Reports Server (NTRS)

Pais, Salvatore Cezar

1999-01-01

The present work reports a study of bubble generation under reduced gravity conditions for both co-flow and cross-flow configurations. Experiments were performed aboard the DC-9 Reduced Gravity Aircraft at NASA Glenn Research Center, using an air-water system. Three different flow tube diameters were used: 1.27, 1.9, and 2.54 cm. Two different ratios of air injection nozzle to tube diameters were considered: 0.1 and 0.2. Gas and liquid volumetric flow rates were varied from 10 to 200 ml/s. It was experimentally observed that with increasing superficial liquid velocity, the bubbles generated decreased in size. The bubble diameter was shown to increase with increasing air injection nozzle diameters. As the tube diameter was increased, the size of the detached bubbles increased. Likewise, as the superficial liquid velocity was increased, the frequency of bubble formation increased and thus the time to detach forming bubbles decreased. Independent of the flow configuration (for either single nozzle or multiple nozzle gas injection), void fraction and hence flow regime transition can be controlled in a somewhat precise manner by solely varying the gas and liquid volumetric flow rates. On the other hand, it is observed that uniformity of bubble size can be controlled more accurately by using single nozzle gas injection than by using multiple port injection, since this latter system gives rise to unpredictable coalescence of adjacent bubbles. A theoretical model, based on an overall force balance, is employed to study single bubble generation in the dynamic and bubbly flow regime. Under conditions of reduced gravity, the gas momentum flux enhances bubble detachment; however, the surface tension forces at the nozzle tip inhibits bubble detachment. Liquid drag and inertia can act either as attaching or detaching force, depending on the relative velocity of the bubble with respect to the surrounding liquid. Predictions of the theoretical model compare well with performed experiments. However, at higher superficial,liquid velocities, the bubble neck length begins to significantly deviate from the value of the air injection nozzle diameter and thus the theory no longer predicts the experiment behavior. Effects of fluid properties, injection geometry and flow conditions on generated bubble size are investigated using the theoretical model. It is shown that bubble diameter is larger in a reduced gravity environment than in a normal gravity environment at similar flow condition and flow geometry.

Exploring the relationship between work-related rumination, sleep quality, and work-related fatigue.

PubMed

Querstret, Dawn; Cropley, Mark

2012-07-01

This study examined the association among three conceptualizations of work-related rumination (affective rumination, problem-solving pondering, and detachment) with sleep quality and work-related fatigue. It was hypothesized that affective rumination and poor sleep quality would be associated with increased fatigue and that problem-solving pondering and detachment would be associated with decreased fatigue. The mediating effect of sleep quality on the relationship between work-related rumination and fatigue was also tested. An online questionnaire was completed by a heterogeneous sample of 719 adult workers in diverse occupations. The following variables were entered as predictors in a regression model: affective rumination, problem-solving pondering, detachment, and sleep quality. The dependent variables were chronic work-related fatigue (CF) and acute work-related fatigue (AF). Affective rumination was the strongest predictor of increased CF and AF. Problem-solving pondering was a significant predictor of decreased CF and AF. Poor sleep quality was predictive of increased CF and AF. Detachment was significantly negatively predictive for AF. Sleep quality partially mediated the relationship between affective rumination and fatigue and between problem-solving pondering and fatigue. Work-related affective rumination appears more detrimental to an individual's ability to recover from work than problem-solving pondering. In the context of identifying mechanisms by which demands at work are translated into ill-health, this appears to be a key finding and suggests that it is the type of work-related rumination, not rumination per se, that is important.

In vitro model to study the effects of matrix stiffening on Ca2+ handling and myofilament function in isolated adult rat cardiomyocytes.

PubMed

van Deel, Elza D; Najafi, Aref; Fontoura, Dulce; Valent, Erik; Goebel, Max; Kardux, Kim; Falcão-Pires, Inês; van der Velden, Jolanda

2017-07-15

This paper describes a novel model that allows exploration of matrix-induced cardiomyocyte adaptations independent of the passive effect of matrix rigidity on cardiomyocyte function. Detachment of adult cardiomyocytes from the matrix enables the study of matrix effects on cell shortening, Ca 2+ handling and myofilament function. Cell shortening and Ca 2+ handling are altered in cardiomyocytes cultured for 24 h on a stiff matrix. Matrix stiffness-impaired cardiomyocyte contractility is reversed upon normalization of extracellular stiffness. Matrix stiffness-induced reduction in unloaded shortening is more pronounced in cardiomyocytes isolated from obese ZSF1 rats with heart failure with preserved ejection fraction compared to lean ZSF1 rats. Extracellular matrix (ECM) stiffening is a key element of cardiac disease. Increased rigidity of the ECM passively inhibits cardiac contraction, but if and how matrix stiffening also actively alters cardiomyocyte contractility is incompletely understood. In vitro models designed to study cardiomyocyte-matrix interaction lack the possibility to separate passive inhibition by a stiff matrix from active matrix-induced alterations of cardiomyocyte properties. Here we introduce a novel experimental model that allows exploration of cardiomyocyte functional alterations in response to matrix stiffening. Adult rat cardiomyocytes were cultured for 24 h on matrices of tuneable stiffness representing the healthy and the diseased heart and detached from their matrix before functional measurements. We demonstrate that matrix stiffening, independent of passive inhibition, reduces cell shortening and Ca 2+ handling but does not alter myofilament-generated force. Additionally, detachment of adult cultured cardiomyocytes allowed the transfer of cells from one matrix to another. This revealed that stiffness-induced cardiomyocyte changes are reversed when matrix stiffness is normalized. These matrix stiffness-induced changes in cardiomyocyte function could not be explained by adaptation in the microtubules. Additionally, cardiomyocytes isolated from stiff hearts of the obese ZSF1 rat model of heart failure with preserved ejection fraction show more pronounced reduction in unloaded shortening in response to matrix stiffening. Taken together, we introduce a method that allows evaluation of the influence of ECM properties on cardiomyocyte function separate from the passive inhibitory component of a stiff matrix. As such, it adds an important and physiologically relevant tool to investigate the functional consequences of cardiomyocyte-matrix interactions. © 2017 The Authors. The Journal of Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society.

An Assessment of Molten Metal Detachment Hazards for Electron Beam Welding in the Space Environment: Analysis and Test Results

NASA Technical Reports Server (NTRS)

Nunes, A. C., Jr.; Russell, C.; Bhat, B.; Fragomeni, J. M.

1998-01-01

Conditions under which molten metal detachments might occur in a space welding environment are analyzed. A weld pool detachment parameter specifying conditions for pool detachment by impact is derived and corroborated by experimental evidence. Impact detachment for the pool is unlikely. Impact detachment for a drop of metal on the end of the weld wire may be possible under extreme conditions. Other potential causes of molten metal detachment considered, vaporization pressure forces and wire flickout from the pool, did not appear to present significant detachment threats.

Curcumin targets breast cancer stem-like cells with microtentacles that persist in mammospheres and promote reattachment.

PubMed

Charpentier, Monica S; Whipple, Rebecca A; Vitolo, Michele I; Boggs, Amanda E; Slovic, Jana; Thompson, Keyata N; Bhandary, Lekhana; Martin, Stuart S

2014-02-15

Cancer stem-like cells (CSC) and circulating tumor cells (CTC) have related properties associated with distant metastasis, but the mechanisms through which CSCs promote metastasis are unclear. In this study, we report that breast cancer cell lines with more stem-like properties display higher levels of microtentacles (McTN), a type of tubulin-based protrusion of the plasma cell membrane that forms on detached or suspended cells and aid in cell reattachment. We hypothesized that CSCs with large numbers of McTNs would more efficiently attach to distant tissues, promoting metastatic efficiency. The naturally occurring stem-like subpopulation of the human mammary epithelial (HMLE) cell line presents increased McTNs compared with its isogenic non-stem-like subpopulation. This increase was supported by elevated α-tubulin detyrosination and vimentin protein levels and organization. Increased McTNs in stem-like HMLEs promoted a faster initial reattachment of suspended cells that was inhibited by the tubulin-directed drug, colchicine, confirming a functional role for McTNs in stem cell reattachment. Moreover, live-cell confocal microscopy showed that McTNs persist in breast stem cell mammospheres as flexible, motile protrusions on the surface of the mammosphere. Although exposed to the environment, they also function as extensions between adjacent cells along cell-cell junctions. We found that treatment with the breast CSC-targeting compound curcumin rapidly extinguished McTN in breast CSC, preventing reattachment from suspension. Together, our results support a model in which breast CSCs with cytoskeletal alterations that promote McTNs can mediate attachment and metastasis but might be targeted by curcumin as an antimetastatic strategy. ©2013 AACR.

Curcumin targets breast cancer stem-like cells with microtentacles that persist in mammospheres and promote reattachment

PubMed Central

Charpentier, Monica S.; Whipple, Rebecca A.; Vitolo, Michele I.; Boggs, Amanda E.; Slovic, Jana; Thompson, Keyata N.; Bhandary, Lekhana; Martin, Stuart S.

2014-01-01

Cancer stem-like cells (CSC) and circulating tumor cells (CTCs) have related properties associated with distant metastasis, but the mechanisms through which CSCs promote metastasis are unclear. In this study, we report that breast cancer cell lines with more stem-like properties display higher levels of microtentacles (McTNs), a type of tubulin-based protrusion of the plasma cell membrane which forms on detached or suspended cells and aid in cell reattachment. We hypothesized that CSCs with large numbers of McTNs would more efficiently attach to distant tissues, promoting metastatic efficiency. The naturally occurring stem-like subpopulation of the HMLE breast cell line presents increased McTNs compared to its isogenic non-stem-like subpopulation. This increase was supported by elevated α-tubulin detyrosination and vimentin protein levels and organization. Increased McTNs in stem-like HMLEs promoted a faster initial reattachment of suspended cells that was inhibited by the tubulin-directed drug, colchicine, confirming a functional role for McTN in stem cell reattachment. Moreover, live cell confocal microscopy showed that McTN persist in breast stem cell mammospheres as flexible, motile protrusions on the surface of the mammosphere. While exposed to the environment, they also function as extensions between adjacent cells along cell-cell junctions. We found that treatment with the breast CSC-targeting compound curcumin rapidly extinguished McTN in breast CSC, preventing reattachment from suspension. Together, our results support a model in which breast CSCs with cytoskeletal alterations that promote McTN can mediate attachment and metastasis but might be targeted by curcumin as an anti-metastatic strategy. PMID:24371229

Rapid detachment of Giardia lamblia trophozoites as a mechanism of antimicrobial action of the isoflavone formononetin

PubMed Central

Lauwaet, Tineke; Andersen, Yolanda; Van de Ven, Liesbeth; Eckmann, Lars; Gillin, Frances D.

2010-01-01

Objectives Attachment to the small intestinal mucosa is crucial for initiating and maintaining Giardia infection. We tested the effect of isoflavones on Giardia attachment. Methods We evaluated the effect of formononetin on trophozoite attachment to glass, to intestinal epithelial cell layers in vitro and to murine small intestinal explants, and on the intestinal load in mice. Results We found that the isoflavone formononetin inhibits both attachment and flagellar motility within minutes and reduces the trophozoite load of Giardia in mice within 1.5 h after treatment. Conclusions The antigiardial activity of formononetin is at least partially due to its capacity to rapidly detach trophozoites. PMID:20067984

Biofilm regeneration on carriers in MBBR used for vitamin C wastewater treatment.

PubMed

Hu, X B; Wang, Zh; Xu, K; Ren, H Q

2013-01-01

Methods were investigated for biofilm regeneration on carriers in a moving bed biofilm reactor used for vitamin C production wastewater treatment. Three ordinary chemical cleaning agents (hydrochloric acid, sodium hydroxide, and sodium hypochlorite) and physical drying were chosen for evaluation as methods for biofilm detachment. The results showed that these methods all had some degree of biofilm removal effectiveness. Treatment with 3% hydrochloric acid (w/w) achieved the maximum degree of biofilm detachment, at 75.2%. Biofilm biomass re-formed on carriers, from the maximum degree of biofilm detachment, quickly by an increase of 76.17 g m(-2) month(-1). It was concluded that treatment with 3% acid was the best choice for biofilm removal and regeneration.

PKCδ Regulates Force Signaling during VEGF/CXCL4 Induced Dissociation of Endothelial Tubes

PubMed Central

Jamison, Joshua; Wang, James H-C.; Wells, Alan

2014-01-01

Wound healing requires the vasculature to re-establish itself from the severed ends; endothelial cells within capillaries must detach from neighboring cells before they can migrate into the nascent wound bed to initiate angiogenesis. The dissociation of these endothelial capillaries is driven partially by platelets' release of growth factors and cytokines, particularly the chemokine CXCL4/platelet factor-4 (PF4) that increases cell-cell de-adherence. As this retraction is partly mediated by increased transcellular contractility, the protein kinase c-δ/myosin light chain-2 (PKCδ/MLC-2) signaling axis becomes a candidate mechanism to drive endothelial dissociation. We hypothesize that PKCδ activation induces contractility through MLC-2 to promote dissociation of endothelial cords after exposure to platelet-released CXCL4 and VEGF. To investigate this mechanism of contractility, endothelial cells were allowed to form cords following CXCL4 addition to perpetuate cord dissociation. In this study, CXCL4-induced dissociation was reduced by a VEGFR inhibitor (sunitinib malate) and/or PKCδ inhibition. During combined CXCL4+VEGF treatment, increased contractility mediated by MLC-2 that is dependent on PKCδ regulation. As cellular force is transmitted to focal adhesions, zyxin, a focal adhesion protein that is mechano-responsive, was upregulated after PKCδ inhibition. This study suggests that growth factor regulation of PKCδ may be involved in CXCL4-mediated dissociation of endothelial cords. PMID:24699667

PKCδ regulates force signaling during VEGF/CXCL4 induced dissociation of endothelial tubes.

PubMed

Jamison, Joshua; Wang, James H-C; Wells, Alan

2014-01-01

Wound healing requires the vasculature to re-establish itself from the severed ends; endothelial cells within capillaries must detach from neighboring cells before they can migrate into the nascent wound bed to initiate angiogenesis. The dissociation of these endothelial capillaries is driven partially by platelets' release of growth factors and cytokines, particularly the chemokine CXCL4/platelet factor-4 (PF4) that increases cell-cell de-adherence. As this retraction is partly mediated by increased transcellular contractility, the protein kinase c-δ/myosin light chain-2 (PKCδ/MLC-2) signaling axis becomes a candidate mechanism to drive endothelial dissociation. We hypothesize that PKCδ activation induces contractility through MLC-2 to promote dissociation of endothelial cords after exposure to platelet-released CXCL4 and VEGF. To investigate this mechanism of contractility, endothelial cells were allowed to form cords following CXCL4 addition to perpetuate cord dissociation. In this study, CXCL4-induced dissociation was reduced by a VEGFR inhibitor (sunitinib malate) and/or PKCδ inhibition. During combined CXCL4+VEGF treatment, increased contractility mediated by MLC-2 that is dependent on PKCδ regulation. As cellular force is transmitted to focal adhesions, zyxin, a focal adhesion protein that is mechano-responsive, was upregulated after PKCδ inhibition. This study suggests that growth factor regulation of PKCδ may be involved in CXCL4-mediated dissociation of endothelial cords.

Upstream Density for Plasma Detachment with Conventional and Lithium Vapor-Box Divertors

NASA Astrophysics Data System (ADS)

Goldston, Rj; Schwartz, Ja

2016-10-01

Fusion power plants are likely to require detachment of the divertor plasma from material targets. The lithium vapor box divertor is designed to achieve this, while limiting the flux of lithium vapor to the main plasma. We develop a simple model of near-detachment to evaluate the required upstream plasma density, for both conventional and lithium vapor-box divertors, based on particle and dynamic pressure balance between up- and down-stream, at near-detachment conditions. A remarkable general result is found, not just for lithium-induced detachment, that the upstream density divided by the Greenwald-limit density scales as (P 5 / 8 /B 3 / 8) Tdet1 / 2 / (ɛcool + γTdet) , with no explicit size scaling. Tdet is the temperature just before strong pressure loss, 1/2 of the ionization potential of the dominant recycling species, ɛcool is the average plasma energy lost per injected hydrogenic and impurity atom, and γ is the sheath heat transmission factor. A recent 1-D calculation agrees well with this scaling. The implication is that the plasma exhaust problem cannot be solved by increasing R. Instead significant innovation, such as the lithium vapor box divertor, will be required. This work supported by DOE Contract No. DE-AC02-09CH11466.

Limbic and prefrontal responses to facial emotion expressions in depersonalization.

PubMed

Lemche, Erwin; Surguladze, Simon A; Giampietro, Vincent P; Anilkumar, Ananthapadmanabha; Brammer, Michael J; Sierra, Mauricio; Chitnis, Xavier; Williams, Steven C R; Gasston, David; Joraschky, Peter; David, Anthony S; Phillips, Mary L

2007-03-26

Depersonalization disorder, characterized by emotional detachment, has been associated with increased prefrontal cortical and decreased autonomic activity to emotional stimuli. Event-related fMRI with simultaneous measurements of skin conductance levels occurred in nine depersonalization disorder patients and 12 normal controls to neutral, mild and intense happy and sad facial expressions. Patients, but not controls, showed decreases in subcortical limbic activity to increasingly intense happy and sad facial expressions, respectively. For both happy and sad expressions, negative correlations between skin conductance measures in bilateral dorsal prefrontal cortices occurred only in depersonalization disorder patients. Abnormal decreases in limbic activity to increasingly intense emotional expressions, and increases in dorsal prefrontal cortical activity to emotionally arousing stimuli may underlie the emotional detachment of depersonalization disorder.

Plasma detachment in divertor tokamaks

NASA Astrophysics Data System (ADS)

Leonard, A. W.

2018-04-01

Observations of divertor plasma detachment in tokamaks are reviewed. Plasma detachment is characterized in terms of transport and dissipation of power, momentum and particle flux along the open field lines from the midplane to the divertor. Asymmetries in detachment onset and other characteristics between the inboard and outboard divertor plasmas is found to be primarily driven by plasma E× B drifts. The effect of divertor plate geometry and magnetic configuration on divertor detachment is summarized. Control of divertor detachment has progressed with a development of a number of diagnostics to characterize the detached state in real-time. Finally the compatibility of detached divertor operation with high performance core plasmas is examined.

Meta-Analysis of Postoperative Outcome Parameters Comparing Descemet Membrane Endothelial Keratoplasty Versus Descemet Stripping Automated Endothelial Keratoplasty.

PubMed

Pavlovic, Ivana; Shajari, Mehdi; Herrmann, Eva; Schmack, Ingo; Lencova, Anna; Kohnen, Thomas

2017-12-01

This meta-analysis compares Descemet membrane endothelial keratoplasty (DMEK) with Descemet stripping automated endothelial keratoplasty (DSAEK) to evaluate their strength and weakness profiles. Meta-analysis. We performed a meta-analysis and searched the peer-reviewed literature in PubMed, Cochrane Library, Web of Science, and Embase following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Prospective and retrospective trials performing and comparing DMEK and DSAEK were included. Effects were calculated as odds ratios or standardized mean differences. A total of 11 studies with a total of 723 eyes (350 DMEK and 373 DSAEK) were included. No significant difference was found regarding the total detachment rate, graft failure, and rejection. One of 340 eyes undergoing DMEK showed total detachment and 5 of 363 eyes undergoing DSAEK showed total detachment (P = 0.28). Six of 280 eyes undergoing DMEK showed graft failure; 1 of 313 eyes undergoing DSAEK developed this complication (P = 0.18). No rejection was observed in 158 eyes undergoing DMEK; 4 cases of rejection occurred in 196 eyes undergoing DSAEK (P = 0.37). No significant difference was found regarding endothelial cell loss (P = 0.48). There was a significantly higher partial detachment rate with DMEK: 88 of 340 eyes undergoing DMEK showed partial detachment; 16 of 363 eyes undergoing DSAEK showed this complication (P < 0.00001). DMEK was superior in best-corrected visual acuity after 6 months (P < 0.001), subjective evaluation of visual acuity (P = 0.001), patient satisfaction (P < 0.001), and was the method preferred by patients (P = 0.001). DMEK and DSAEK have a similar complication profile. However, the superiority in the visual outcome and patient satisfaction makes DMEK the preferred option for most patients.

GRAFT-VERSUS-HOST DISEASE PANUVEITIS AND BILATERAL SEROUS DETACHMENTS: MULTIMODAL IMAGING ANALYSIS.

PubMed

Jung, Jesse J; Chen, Michael H; Rofagha, Soraya; Lee, Scott S

2017-01-01

To report the multimodal imaging findings and follow-up of a case of graft-versus-host disease-induced bilateral panuveitis and serous retinal detachments after allogenic bone marrow transplant for acute myeloid leukemia. A 75-year-old black man presented with acute decreased vision in both eyes for 1 week. Clinical examination and multimodal imaging, including spectral domain optical coherence tomography, fundus autofluorescence, fluorescein angiography, and swept-source optical coherence tomography angiography (Investigational Device; Carl Zeiss Meditec Inc) were performed. Clinical examination of the patient revealed anterior and posterior inflammation and bilateral serous retinal detachments. Ultra-widefield fundus autofluorescence demonstrated hyperautofluorescence secondary to subretinal fluid; and fluorescein angiography revealed multiple areas of punctate hyperfluorescence, leakage, and staining of the optic discs. Spectral domain and enhanced depth imaging optical coherence tomography demonstrated subretinal fluid, a thickened, undulating retinal pigment epithelium layer, and a thickened choroid in both eyes. En-face swept-source optical coherence tomography angiography did not show any retinal vascular abnormalities but did demonstrate patchy areas of decreased choriocapillaris flow. An extensive systemic infectious and malignancy workup was negative and the patient was treated with high-dose oral prednisone immunosuppression. Subsequent 6-month follow-up demonstrated complete resolution of the inflammation and bilateral serous detachments after completion of the prednisone taper over a 3-month period. Graft-versus-host disease panuveitis and bilateral serous retinal detachments are rare complications of allogenic bone marrow transplant for acute myeloid leukemia and can be diagnosed with clinical and multimodal imaging analysis. This form of autoimmune inflammation may occur after the recovery of T-cell activity within the donor graft targeting the host. Infectious and recurrent malignancy must be ruled out before initiation of immunosuppression, which can affectively treat this form of graft-versus-host disease.

Promotion of malignant phenotype after disruption of the three-dimensional structure of cultured spheroids from colorectal cancer.

PubMed

Piulats, Jose M; Kondo, Jumpei; Endo, Hiroko; Ono, Hiromasa; Hagihara, Takeshi; Okuyama, Hiroaki; Nishizawa, Yasuko; Tomita, Yasuhiko; Ohue, Masayuki; Okita, Kouki; Oyama, Hidejiro; Bono, Hidemasa; Masuko, Takashi; Inoue, Masahiro

2018-03-23

Individual and small clusters of cancer cells may detach from the edges of a main tumor and invade vessels, which can act as the origin of metastasis; however, the mechanism for this phenomenon is not well understood. Using cancer tissue-originated spheroids, we studied whether disturbing the 3D architecture of cancer spheroids can provoke the reformation process and progression of malignancy. We developed a mechanical disruption method to achieve homogenous disruption of the spheroids while maintaining cell-cell contact. After the disruption, 9 spheroid lines from 9 patient samples reformed within a few hours, and 3 of the 9 lines exhibited accelerated spheroid growth. Marker expression, spheroid forming capacity, and tumorigenesis indicated that stemness increased after spheroid disruption. In addition, the spheroid forming capacity increased in 6 of 11 spheroid lines. The disruption signature determined by gene expression profiling supported the incidence of remodeling and predicted the prognosis of patients with colorectal cancer. Furthermore, WNT and HER3 signaling were increased in the reformed spheroids, and suppression of these signaling pathways attenuated the increased proliferation and stemness after the disruption. Overall, the disruption and subsequent reformation of cancer spheroids promoted malignancy-related phenotypes through the activation of the WNT and ERBB pathways.

Hydraulic parameters in eroding rills and their influence on detachment processes

NASA Astrophysics Data System (ADS)

Wirtz, Stefan; Seeger, Manuel; Zell, Andreas; Wagner, Christian; Wengel, René; Ries, Johannes B.

2010-05-01

In many experiments as well in laboratory as in field experiments the correlations between the detachment rate and different hydraulic parameters are calculated. The used parameters are water depth, runoff, shear stress, unit length shear force, stream power, Reynolds- and Froude number. The investigations show even contradictory results. In most soil erosion models like the WEPP model, the shear stress is used to predict soil detachment rates. But in none of the WEPP datasets, the shear stress showed the best correlation to the detachment rate. In this poster we present the results of several rill experiments in Andalusia from 2008 and 2009. With the used method, it is possible to measure the needed factors to calculate the mentioned parameters. Water depth is measured by an ultrasonic sensor, the runoff values are calculated by combining flow velocity and flow diameter. The parameters wetted perimeter, flow diameter and hydraulic radius can be calculated from the measured rill cross sections and the measured water levels. In the sample density values, needed for calculation of shear stress, unit length shear force and stream power, the sediment concentration and the grain density are are considered. The viscosity of the samples was measured with a rheometer. The result of this measurements shows, that there is a very high linear correlation (R² = 0.92) between sediment concentration and the dynamic viscosity. The viscosity seems to be an important factor but it is only used in the Reynolds-number-equation, in other equations it is neglected. But the viscosity value increases with increasing sediment concentration and hence the influence also increases and the in multiclications negiligible viscosity value of 1 only counts for clear water. The correlations between shear stress, unit length shear force and stream power at the x-axis and the detachment rate at the ordinate show, that there is not one fixed parameter that always displays the best correlation to the detachment rate. The best hit does not change from one experiment to another, it changes from one measuring point to another. Different processes in rill erosion are responsible for the changing correlations. In some cases no one of the parameters shows an acceptable correlation to the soil detachment, because these factors describe fluvial processes. Our experiments show, that not the fluvial processes cause the main sediment procduction in the rills, but bank failure or knickpoint and headcut retreat and these processes are more gravitative than fluvial. Another sediment producing process is the abrupt spill over of plunge pools, a process not realy fluvial and not realy gravitativ. In some experiments, the highest sediment concentrations were measured at the slowly flowing waterfront that only transports the loose material. But all these processes are not considered in soil erosion models. Hence, hydraulic parameters alone are not sufficient to predict detachment rates. They cover the fluvial incising in the rill's bottom, but the main sediment sources are not considered satisying in its equations.

Coupling between apical tension and basal adhesion allow epithelia to collectively sense and respond to substrate topography over long distances.

PubMed

Broaders, Kyle E; Cerchiari, Alec E; Gartner, Zev J

2015-12-01

Epithelial sheets fold into complex topographies that contribute to their function in vivo. Cells can sense and respond to substrate topography in their immediate vicinity by modulating their interfacial mechanics, but the extent to which these mechanical properties contribute to their ability to sense substrate topography across length scales larger than a single cell has not been explored in detail. To study the relationship between the interfacial mechanics of single cells and their collective behavior as tissues, we grew cell-sheets on substrates engraved with surface features spanning macroscopic length-scales. We found that many epithelial cell-types sense and respond to substrate topography, even when it is locally nearly planar. Cells clear or detach from regions of local negative curvature, but not from regions with positive or no curvature. We investigated this phenomenon using a finite element model where substrate topography is coupled to epithelial response through a balance of tissue contractility and adhesive forces. The model correctly predicts the focal sites of cell-clearing and epithelial detachment. Furthermore, the model predicts that local tissue response to substrate curvature is a function of the surrounding topography of the substrate across long distances. Analysis of cell-cell and cell-substrate contact angles suggests a relationship between these single-cell interfacial properties, epithelial interfacial properties, and collective epithelial response to substrate topography. Finally, we show that contact angles change upon activation of oncogenes or inhibition of cell-contractility, and that these changes correlate with collective epithelial response. Our results demonstrate that in mechanically integrated epithelial sheets, cell contractility can be transmitted through multiple cells and focused by substrate topography to affect a behavioral response at distant sites.

KSHV MicroRNAs Repress Tropomyosin 1 and Increase Anchorage-Independent Growth and Endothelial Tube Formation

PubMed Central

Kieffer-Kwon, Philippe; Happel, Christine; Uldrick, Thomas S.; Ramalingam, Dhivya; Ziegelbauer, Joseph M.

2015-01-01

Kaposi’s sarcoma (KS) is characterized by highly vascularized spindle-cell tumors induced after infection of endothelial cells by Kaposi’s sarcoma-associated herpesvirus (KSHV). In KS tumors, KSHV expresses only a few latent proteins together with 12 pre-microRNAs. Previous microarray and proteomic studies predicted that multiple splice variants of the tumor suppressor protein tropomyosin 1 (TPM1) were targets of KSHV microRNAs. Here we show that at least two microRNAs of KSHV, miR-K2 and miR-K5, repress protein levels of specific isoforms of TPM1. We identified a functional miR-K5 binding site in the 3’ untranslated region (UTR) of one TPM1 isoform. Furthermore, the inhibition or loss of miR-K2 or miR-K5 restores expression of TPM1 in KSHV-infected cells. TPM1 protein levels were also repressed in KSHV-infected clinical samples compared to uninfected samples. Functionally, miR-K2 increases viability of unanchored human umbilical vein endothelial cells (HUVEC) by inhibiting anoikis (apoptosis after cell detachment), enhances tube formation of HUVECs, and enhances VEGFA expression. Taken together, KSHV miR-K2 and miR-K5 may facilitate KSHV pathogenesis. PMID:26263384

[Treatment delay in patients with first episode of retinal detachment in the studied eye in Midi-Pyrénées].

PubMed

Tolou, C; Mahieu-Durringer, L; Cassagne, M; Hamid, S; Billette de Villemeur, R; Gualino, V; Susini, A; Pagot Mathis, V; Gallini, A; Soler, V

2016-01-01

Retinal detachment (RD) is a potentially blinding condition. Delay in management is a major prognostic factor. In our study, we analyzed the treatment delay for retinal detachments in the Midi-Pyrenees area, and factors which may influence it. Observational, cross-sectional, multicentric study, carried out over a 6-month period. time between diagnosis and surgery. Secondary outcome: time between first symptoms and surgery. Non-parametric tests were used to analyze the influence of sociodemographic features, clinical features, distance between home and surgical center, and occurrence over a weekend. One hundred and fiftty-nine patients were included. The mean time between diagnosis and surgery was 4.4 ± 12.3 days (2.7 ± 4.3 for recent RD, less than 1 month), and was increased by the presence of a weekend (P<0.001), or of a weekend with public holiday (P=0.023), and by macular detachment (P=0.008). The mean time between first symptoms and surgery was 12.0 days and was increased by the absence of RD history (P=0.023), and by macular detachment (P=0.046). No association was observed between these times to surgery and the distance between the patient's home address and the place of surgery. The time between diagnosis and surgery was relatively short in the Midi-Pyrénées area, but we often noted a delayed diagnosis, which may be due to the patient's lack of awareness of the symptoms and difficult access to specialty consultations. However, no relationship was found between this time-to-surgery and the distance between the patient's home and the surgical center. Copyright © 2015. Published by Elsevier Masson SAS.

Rate and Localization of Graft Detachment in Descemet Membrane Endothelial Keratoplasty.

PubMed

Maier, Anna-Karina B; Gundlach, Enken; Pilger, Daniel; Rübsam, Anne; Klamann, Matthias K J; Gonnermann, Johannes; Bertelmann, Eckart; Joussen, Antonia M; Torun, Necip

2016-03-01

To investigate the rate and localization of graft detachment after Descemet membrane endothelial keratoplasty. Sixty-six consecutive cases operated between June and August 2014 at the Charité-Universitätsmedizin Berlin were examined prospectively 1 week postoperatively. A single masked observer analyzed the rate and localization of graft detachment using optical coherence tomography (OCT), and the rebubbling rate was measured. Localization of graft detachment was correlated to the incision approach. Preoperative data were correlated to the rate of graft detachment and rebubbling. Graft detachment occurred in more than 2 clock hours and with postoperative corneal edema in 33.3% and required rebubbling. In 33.3%, graft detachment occurred in more than 2 clock hours and with postoperative corneal edema and required rebubbling. The mean graft detachment rate was 8.3% per clock hour. A significantly higher graft detachment rate was noted in the inferior clock hours (21.1%, P < 0.0001, 16.7%, P = 0.003). Only higher age of the patient correlated to a higher rate of graft detachment (P = 0.022). No correlation was found between localization of graft detachment and the incision approach (P = 0.615). The graft detachment rate is high after Descemet membrane endothelial keratoplasty, but detachment is usually peripheral, partial and mainly inferior and involves only a few clock hours. Only higher age of the patient is strongly associated with a higher rate of graft detachment. The incision approach is not significantly correlated with the localization of graft detachment. Therefore, the postoperative supine position of the patient seems to be of major importance.Clinical Trial Registration-URL: http://www.clinicaltrials.gov. Unique identifier: NCT02020044.

Role of Outer Membrane C-Type Cytochromes MtrC and OmcA in Shewanella Oneidensis MR-1 Cell Production, Accumulation, and Detachment During Respiration on Hematite

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

Mitchell, Andrew C.; Peterson, L.; Reardon, Catherine L.

2012-07-01

Solid phase iron oxides are considered to be important terminal electron acceptors for microbial respiration in many anoxic environments. Besides the knowledge that cells attach to and reduce these substrates, other aspects of surface-associated cell behavior and the related cell surface components that influence cell-mineral interactions are not well understood. In the present study, wild-type cells of the dissimilatory iron-reducing bacterium Shewanella oneidensis MR-1 formed thin biofilms one-to-two cell layers in thickness when respiring on natural specular hematite under flow conditions similar to those which exist in aquatic sediments and subsurface environments. The distribution of cells within the biofilm indicatedmore » that direct contact was not required for electron transfer from cells to the mineral surface. Detached biomass in the form of single cells represented >99% of the surface-associated wild-type cell production from respiration on hematite over the biofilm life cycle. A mutant deficient in the outer membrane c35 type cytochrome OmcA, while still able to respire and replicate on hematite, established a lower steady-state cell density on the mineral surface than that of the wild-type strain. A mutant deficient in MtrC, another outer membrane c-type cytochrome, and a mutant deficient in both cytochromes were unable to reduce sufficient amounts of hematite to support detectable growth on the mineral surface. When considered in the context of previous work, the results support a growing body of evidence that the relative importance of OmcA and MtrC to cell respiration and replication depends on the form of iron oxide available as terminal electron acceptor.« less

Plasma detachment in divertor tokamaks

DOE PAGES

Leonard, A. W.

2018-02-07

In this study, observations of divertor plasma detachment in tokamaks are reviewed. Plasma detachment is characterized in terms of transport and dissipation of power, momentum and particle flux along the open field lines from the midplane to the divertor. Asymmetries in detachment onset and other characteristics between the inboard and outboard divertor plasmas is found to be primarily driven by plasmamore » $$\\vec{E}$$ x $$\\vec{B}$$ drifts. The effect of divertor plate geometry and magnetic configuration on divertor detachment is summarized. Control of divertor detachment has progressed with a development of a number of diagnostics to characterize the detached state in real-time. Finally the compatibility of detached divertor operation with high performance core plasmas is examined.« less

Plasma detachment in divertor tokamaks

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

Leonard, A. W.

In this study, observations of divertor plasma detachment in tokamaks are reviewed. Plasma detachment is characterized in terms of transport and dissipation of power, momentum and particle flux along the open field lines from the midplane to the divertor. Asymmetries in detachment onset and other characteristics between the inboard and outboard divertor plasmas is found to be primarily driven by plasmamore » $$\\vec{E}$$ x $$\\vec{B}$$ drifts. The effect of divertor plate geometry and magnetic configuration on divertor detachment is summarized. Control of divertor detachment has progressed with a development of a number of diagnostics to characterize the detached state in real-time. Finally the compatibility of detached divertor operation with high performance core plasmas is examined.« less

The Detached Haze Layer in Titan's Mesosphere

NASA Astrophysics Data System (ADS)

Lavvas, P.; Yelle, R. V.; Vuitton, V.

2008-12-01

The Cassini observations reveal the presence of a detached haze layer in Titan's mesosphere at an altitude of 520 km, well above the stratosphere. Observations of scattered light made by the Imaging Science Subsystem (ISS) reveal a clearly defined layer encircling low and mid-latitude regions. The aerosol layer is also detected in stellar occultation measurements of UV extinction by the UltraViolet Imaging Spectrometer (UVIS). The haze is a global and permanent feature of Titan's atmosphere. Furthermore the location of the detached haze layer is coincident with and the likely cause of a local maximum in the temperature profile measured by the Huygens Atmospheric Structure Instrument (HASI). This temperature inversion is also permanent and global, having been detected in ground-based stellar occultations. The correlation between the extinction profile and the temperature maximum imply that the detached haze cannot be due to condensation, as previously suggested. Previously, Voyager high phase angle images at 500 nm revealed a detached haze layer near 350 km, more than 150 km lower than the Cassini layer. Close examination of the Voyager images suggests that the Cassini detached layer at 520 km is a separate phenomenon rather than a change in the Voyager detached layer. Analysis of the observed optical properties suggests that the average size of particles in the Cassini detached layer is < 45 nm, with an imaginary index k < 0.3 at 187.5 nm, while Non-LTE calculations of the temperature perturbation induced by the detached haze show that the average particle size must be greater than 35 nm for reproducing the heating rate implied by the HASI temperature profile. Calculation of the sedimentation velocity of the particles, coupled with the derived number density, imply a mass flux of 1.9-3.2 × 10-14 g cm-2 s-1. This is approximately equal to the mass flux required to explain the main haze layer and suggests that the stratospheric haze is formed primarily by sedimentation and coagulation of particles in the detached layer. This is further supported from the particle size range retrieved for the detached layer (35-45 nm), being approximately equal to the radii of ~50 nm for the monomers of the aggregate aerosols in the main haze layer. It follows that aerosols on Titan are formed primarily in the thermosphere, rather than the stratosphere as assumed in many pre-Cassini studies. This is consistent with the detection of negatively charged aerosols in the thermosphere by Cassini/CAPS. These conclusions are supported by microphysical aerosol models that couple the detached haze layer and the main haze layer and extend into the thermosphere. Our calculations suggest that the detached haze layer is due to the transition in the growth of particles from spherical shape to aggregates of fractal structure. The rapid increase in the size of the particles with the onset of fractal growth, in combination with the decrease of their settling velocity, casts them invisible in the transition region. This optical illusion process explains the well-defined and symmetric structure of the detached layer, something difficult to explain under a pure advection scenario. Further investigation of the processes defining the growth of the particles is required in order to understand why the transition takes place at this region and how the particles produced at higher altitudes by high energy radical and ion chemistry, are defining the vertical haze opacity in Titan's atmosphere.

Association of Host Genetic Risk Factors with the Course of Cytomegalovirus Retinitis in Patients Infected with HIV

PubMed Central

Sezgin, Efe; Van Natta, Mark L.; Ahuja, Alka; Lyon, Alice; Srivastava, Sunil; Troyer, Jennifer L.; O’Brien, Stephen J.; Jabs, Douglas A.

2010-01-01

Purpose To evaluate the effects of previously reported host genetics factors that influence cytomegalovirus (CMV) retinitis incidence, progression to AIDS, and efficacy of highly active antiretroviral therapy (HAART) for mortality, retinitis progression, and retinal detachment in patients with CMV retinitis and AIDS in the era of HAART. Design Prospective, multicenter, observational study. Methods Cox proportional hazards model based genetic association tests examined the influence of IL-10R1_S420L, CCR5Δ32, CCR2-V64I, CCR5 P1, and SDF-3`A polymorphisms among patients with mortality, retinitis progression, and retinal detachment. Participants were 203 European American and 117 African American patients with AIDS and CMV retinitis. Results European American patients with the CCR5 +.P1.+ promoter haplotype showed increased risk for mortality (HR=1.83; 95% CI: 1.00–3.40; P=0.05). Although the same haplotype also trended for increased risk for mortality in African American patients, the result was not significant (HR=2.28; 95% CI: 0.93–5.60; P=0.07). However, this haplotype was associated with faster retinitis progression in African Americans (HR=5.22; 95% CI: 1.54–17.71; P=0.007). Increased risk of retinitis progression was also evident for African American patients with the SDF1-3′A variant (HR=3.89; 95% CI: 1.42–10.60; P=0.008). In addition, the SDF1-3′A variant increased the retinal detachment risk in this patient group (HR=3.05; 95% CI: 1.01–9.16; P=0.05). Conclusion Besides overall immune health, host genetic factors influence mortality, retinitis progression, and retinal detachment in patients with AIDS and CMV retinitis that are receiving HAART. PMID:21396623

Detachment of solids and nitrifiers in integrated, fixed-film activated sludge systems.

PubMed

Maas, Carol L A; Parker, Wayne J; Legge, Raymond L

2008-12-01

Despite the importance of detachment to biofilm processes, detachment phenomena are not well understood. In this study, researchers investigated biofilm detachment from free-floating biofilm carriers that were established in an integrated, fixed-film activated sludge (IFAS) installation in Mississauga, Ontario. A method for assessing detachment from biofilm carrier systems was devised, evaluated, and refined during this study. In the absence of substrate, superficial air velocity significantly affected the 24-hour detachment rates of total suspended solids from the carriers. Short-term growth conditions did not appear to significantly affect the rate of detachment of solids and nitrifiers. The measured solids-detachment rates were found to be described by a second order function of biofilm attached growth total solids with a detachment coefficient of 0.006 +/- 0.0008 (g/m x d)(-1).

Intracellular Pressure Dynamics in Blebbing Cells

PubMed Central

Strychalski, Wanda; Guy, Robert D.

2016-01-01

Blebs are pressure-driven protrusions that play an important role in cell migration, particularly in three-dimensional environments. A bleb is initiated when the cytoskeleton detaches from the cell membrane, resulting in the pressure-driven flow of cytosol toward the area of detachment and local expansion of the cell membrane. Recent experiments involving blebbing cells have led to conflicting hypotheses regarding the timescale of intracellular pressure propagation. The interpretation of one set of experiments supports a poroelastic model of the cytoplasm that leads to slow pressure equilibration when compared to the timescale of bleb expansion. A different study concludes that pressure equilibrates faster than the timescale of bleb expansion. To address this discrepancy, a dynamic computational model of the cell was developed that includes mechanics of and the interactions among the cytoplasm, the actin cortex, the cell membrane, and the cytoskeleton. The model results quantify the relationship among cytoplasmic rheology, pressure, and bleb expansion dynamics, and provide a more detailed picture of intracellular pressure dynamics. This study shows the elastic response of the cytoplasm relieves pressure and limits bleb size, and that both permeability and elasticity of the cytoplasm determine bleb expansion time. Our model with a poroelastic cytoplasm shows that pressure disturbances from bleb initiation propagate faster than the timescale of bleb expansion and that pressure equilibrates slower than the timescale of bleb expansion. The multiple timescales in intracellular pressure dynamics explain the apparent discrepancy in the interpretation of experimental results. PMID:26958893

Physicochemical approach to freshwater microalgae harvesting with magnetic particles.

PubMed

Prochazkova, Gita; Podolova, Nikola; Safarik, Ivo; Zachleder, Vilem; Branyik, Tomas

2013-12-01

Magnetic harvesting of microalgal biomass provides an attractive alternative to conventional methods. The approach to this issue has so far been pragmatic, focused mainly on finding cheap magnetic agents in combination with harvestable microalgae species. The aim of this work was to study experimentally and theoretically the mechanisms leading to cell-magnetic agent attachment/detachment using real experiments and predictions made by colloidal adhesion (XDLVO) model. Two types of well defined magnetic beads (MBs) carrying ion exchange functional groups (DEAE - diethylaminoethyl and PEI - polyethylenimine) were studied in connection with microalgae (Chlorella vulgaris). Optimal harvesting efficiencies (>90%) were found for DEAE and PEI MBs, while efficient detachment was achieved only for DEAE MBs (>90%). These findings were in accordance with the predictions by XDLVO model. Simultaneously there was found a discrepancy between the XDLVO prediction and the poor detachment of PEI MBs from microalgal surface. This can be ascribed to an additional interaction (probably covalent bonds) between PEI and algal surface, which the XDLVO model is unable to capture given by its non-covalent nature. Copyright © 2013 Elsevier B.V. All rights reserved.

Anticancer effects elicited by combination of Rubus extract with phthalocyanine photosensitiser on MCF-7 human breast cancer cells.

PubMed

George, Blassan P; Abrahamse, Heidi; Hemmaragala, Nanjundaswamy M

2017-09-01

Photodynamic therapy (PDT) is a novel approach for the treatment of cancer and other related diseases. Breast cancer remains the most common cause of cancer-related death in women. This study was carried out to investigate the photosensitizing capacity of Rubus fairholmianus root acetone extract (RFRA) in vitro. RFRA was coupled with phthalocyanine photosensitizer to enhance the therapeutic properties on MCF-7 breast cancer cells. Comparatively low dose photosensitizer (PS) and Rubus extract have been used for the conjugation as it induces cell death at low doses. The diode laser of wavelength 680 nm and 5, 10 and 15 J/cm2 fluencies have been used for PDT experiments/laser irradiation. MCF-7 cells were exposed to Rubus extract and conjugated Rubus-PS for 24 h and analysed the alterations in cell morphology, proliferation, cytotoxicity and apoptosis induction. The PDT-treated cells displayed substantial features of apoptotic cell death by changes in morphology with a reduction in cell number, development of apoptotic bodies and cell detachment from culture plates. Cellular viability (51.25% for RFRA-PS at 15 J/cm2) and Adenosine 5'-triphosphate (ATP) proliferation of treated cells reduced significantly and the cytotoxicity increased in lactate dehydrogenase (LDH) assay. The Annexin V/PI double staining supports the caspase 3/7 activities by the increased apoptotic cells population and the increased levels of cytochrome c. Our results show that the phototoxic properties of RFRA and photosensitizer may be through the caspase-mediated apoptosis and it can be summarised that Rubus may be a potent anticancer plant with phototoxic effects on breast cancer cells. Copyright © 2017 Elsevier B.V. All rights reserved.

Folding of a detachment and fault - Modified detachment folding along a lateral ramp, southwestern Montana, USA

NASA Astrophysics Data System (ADS)

Schmidt, Christopher; Whisner, S. Christopher; Whisner, Jennifer B.

2014-12-01

The inversion of the Middle Proterozoic Belt sedimentary basin during Late Cretaceous thrusting in Montana produced a large eastwardly-convex salient, the southern boundary of which is a 200 km-long oblique to lateral ramp subtended by a detachment between the Belt rocks and Archean basement. A 10 km-long lateral ramp segment exposes the upper levels of the detachment where hanging wall Belt rocks have moved out over the Paleozoic and Mesozoic section. The hanging wall structure consists of a train of high amplitude, faulted, asymmetrical detachment folds. Initial west-east shortening produced layer parallel shortening fabrics and dominantly strike slip faulting followed by symmetrical detachment folding. 'Lock-up' of movement on the detachment surface produced regional simple shear and caused the detachment folds to become asymmetrical and faulted. Folding of the detachment surface after lock-up modified the easternmost detachment folds further into a southeast-verging, overturned fold pair with a ramp-related fault along the base of the stretched mutual limb.

Mesoporous silica nanoparticles engineered for ultrasound-induced uptake by cancer cells.

PubMed

Paris, Juan L; Manzano, Miguel; Cabañas, M Victoria; Vallet-Regí, María

2018-04-05

A novel smart hierarchical ultrasound-responsive mesoporous silica nanocarrier for cancer therapy is presented here. This dynamic nanosystem has been designed to display different surface characteristics during its journey towards tumor cells. Initially, the anticancer-loaded nanocarriers are shielded with a polyethylene glycol layer. Upon exposure to high frequency ultrasound, the polymer shell detaches from the nanoparticles, exposing a positively charged surface. This favors the internalization in human osteosarcoma cells, where the release of topotecan takes place, drastically enhancing the cytotoxic effect.

Embryonic integument and "molts" in Manduca sexta (Insecta, Lepidoptera).

PubMed

Ziese, Stefanie; Dorn, August

2003-02-01

In Manduca sexta the germ band is formed 12 h post-oviposition (p.o.) (=10% development completed) and is located above the yolk at the egg surface. The cells show a polar organization. They are engaged in the uptake and degradation of yolk globules, pinched off from the yolk cells. This process can be observed in the integumental cells during the first growth phase of the embryo that lasts until "katatrepsis," an embryonic movement that takes place at 40% development completed. At 37% development completed, the ectoderm deposits a thin membrane at its apical surface, the first embryonic membrane, which detaches immediately before katatrepsis. The second period of embryonic growth--from katatrepsis to 84 h p.o. (70% development completed)--starts with the deposition of a second embryonic membrane that is somewhat thicker than the first one and shows a trilaminar, cuticulin-like structure. Whereas the apical cell surface is largely smooth during the deposition of the first embryonic membrane, it forms microvilli during deposition of the second one. At the same time, uptake of formed yolk material ceases and the epidermal cells now contain clusters of mitochondria below the apical surface. Rough endoplasmic reticulum (RER) increases in the perinuclear region. The second embryonic membrane detaches about 63 h p.o. At 69 h p.o., a new generation of microvilli forms and islands of a typical cuticulin layer indicate the onset of the deposition of the larval cuticle. The third growth phase is characterized by a steady increase in the embryo length, the deposition of the larval procuticle, and by cuticular tanning at about 100 h p.o. Beginning at that stage, electron-lucent vesicles aggregate below the epidermal surface and are apparently released below the larval cuticle. Manduca sexta is the first holometabolous insect in which the deposition of embryonic membranes and cuticles has been examined by electron microscopy. In correspondence with hemimetabolous insects, the embryo of M. sexta secretes three covers at approximately the same developmental stage. A marked difference: the second embryonic cover, which in Hemimetabola clearly exhibits a cuticular organization, has instead a membranous, cuticulin-like structure. We see the difference as the result of an evolutionary reductional process promoted by the redundancy of embryonic covers in the egg shell. Embryonic "molts" also occur in noninsect arthropods; their phylogenetical aspects are discussed. Copyright 2002 Wiley-Liss, Inc.

Protein disulfide isomerases in the endoplasmic reticulum promote anchorage-independent growth of breast cancer cells.

PubMed

Wise, Randi; Duhachek-Muggy, Sara; Qi, Yue; Zolkiewski, Michal; Zolkiewska, Anna

2016-06-01

Metastatic breast cancer cells are exposed to stress of detachment from the extracellular matrix (ECM). Cultured breast cancer cells that survive this stress and are capable of anchorage-independent proliferation form mammospheres. The purpose of this study was to explore a link between mammosphere growth, ECM gene expression, and the protein quality control system in the endoplasmic reticulum (ER). We compared the mRNA and protein levels of ER folding factors in SUM159PT and MCF10DCIS.com breast cancer cells grown as mammospheres versus adherent conditions. Publicly available gene expression data for mammospheres formed by primary breast cancer cells and for circulating tumor cells (CTCs) were analyzed to assess the status of ECM/ER folding factor genes in clinically relevant samples. Knock-down of selected protein disulfide isomerase (PDI) family members was performed to examine their roles in SUM159PT mammosphere growth. We found that cells grown as mammospheres had elevated expression of ECM genes and ER folding quality control genes. CTC gene expression data for an index patient indicated that upregulation of ECM and ER folding factor genes occurred at the time of acquired therapy resistance and disease progression. Knock-down of PDI, ERp44, or ERp57, three members of the PDI family with elevated protein levels in mammospheres, in SUM159PT cells partially inhibited the mammosphere growth. Thus, breast cancer cell survival and growth under detachment conditions require enhanced assistance of the ER protein folding machinery. Targeting ER folding factors, in particular members of the PDI family, may improve the therapeutic outcomes in metastatic breast cancer.

The Research of Tectonic Framework and the Fault Activity in Large Detachment Basin System on Northern Margin of South China Sea

NASA Astrophysics Data System (ADS)

Pan, L., Sr.; Ren, J.

2017-12-01

The South China Sea (SCS) is one of the largest marginal sea on southeast Asia continental margin, developed Paleogene extension-rifting continental margin system which is rare in the world and preserving many deformed characterizes of this kind system. With the investigation of the SCS, guiding by the development of tectonics and geo-physics, especially the development of tectonics and the high quality seismic data based on the development of geo-physics, people gradually accept that the northern margin of the SCS has some detachment basin characterizes. After researching the northern margin of the SCS, we come up with lithosphere profiles across the shelf, slope and deep sea basin in the northeast of the SCS to confirm the tectonic style of ocean-continental transition and the property of the detachment fault. Furthermore, we describe the outline of large detachment basins at northern SCS. Based on the large number of high-quality 2D and 3D deep seismic profile(TWT,10s), drilling and logging data, combined with domestic and international relevant researches, using basin dynamics and tectono-stratigraphy theory, techniques and methods of geology and geophysics, qualitative and quantitative, we describe the formation of the detachment basin and calculate the fault activity rate, stretching factor and settlement. According to the research, we propose that there is a giant and complete detachment basin system in the northern SCS and suggest three conclusions. First of all, the detachment basin system can be divided into three domains: proximal domain covering the Yangjiang Sag, Shenhu uplift and part of Shunde Sag, necking zone covering part of the Shunde Sag and Heshan Sag, distal domain covering most part of Heshan Sag. Second, the difference of the stretching factor is observed along the three domains of the detachment basin system. The factor of the proximal domain is the minimum among them. On the other side, the distal domain is the maximum among them. This phenomenon can be concluded as the factor is gradually increasing from the continent to the ocean. Third, the development of detachment basin is episodic which can be divided into two stages approximately: the rifting and thermal subsidence.

Neutrophil autophagy and extracellular DNA traps contribute to airway inflammation in severe asthma.

PubMed

Pham, D L; Ban, G-Y; Kim, S-H; Shin, Y S; Ye, Y-M; Chwae, Y-J; Park, H-S

2017-01-01

Autophagy and neutrophil extracellular DNA traps (NETs) are implicated in asthma; however, their roles in asthma pathogenesis have not been elucidated. We compared autophagy and NET production levels from peripheral blood neutrophils (PBNs) of patients with severe asthma (SA) and non-severe asthma (NSA). Additionally, we investigated the inflammatory effects of NETs on human airway epithelial cells (AECs) and peripheral blood eosinophils (PBEs). Peripheral blood neutrophils from patients with SA (n = 30) and NSA (n = 38) were treated with interleukin (IL)-8 (100 ng/mL). Autophagy (light chain 3-II expression) and NET production levels were evaluated by Western blot, immunofluorescence microscopy, and PicoGreen assay. The effects of NETs on AECs were assessed by investigating cell death, cell detachment, expression of occludin and claudin-1, and IL-8 production; the effects of NETs on PBEs were examined by investigating the activation and release of eosinophil cationic protein (ECP) and eosinophil-derived neurotoxin (EDN). Untreated and IL-8-treated PBNs from the SA group produced higher autophagy and NET levels compared with those from the NSA group (P < 0.01). IL-8 increased autophagy and NET levels in PBNs from the SA group, but not from the NSA group. NET levels were correlated with autophagy levels in PBNs (P < 0.001). IL-8-induced NET production levels negatively were correlated with FEV1/FVC (r = -0.700, P = 0.016). NETs induced cell death, detachment, degradation of occludin and claudin-1, and IL-8 production from AECs. Higher levels of NET-induced ECP and EDN were released from PBEs in SA compared with NSA groups. Neutrophil autophagy and NETs could enhance asthma severity by damaging airway epithelium and triggering inflammatory responses of AECs and PBEs. Modulating neutrophil autophagy and NET production may be a new target therapy for SA. © 2016 John Wiley & Sons Ltd.

Quantitative Analysis of Complex Glioma Cell Migration on Electrospun Polycaprolactone Using Time-Lapse Microscopy

PubMed Central

Johnson, Jed; Nowicki, M. Oskar; Lee, Carol H.; Chiocca, E. Antonio; Viapiano, Mariano S.; Lawler, Sean E.

2009-01-01

Malignant gliomas are the most common tumors originating within the central nervous system and account for over 15,000 deaths annually in the United States. The median survival for glioblastoma, the most common and aggressive of these tumors, is only 14 months. Therapeutic strategies targeting glioma cells migrating away from the tumor core are currently hampered by the difficulty of reproducing migration in the neural parenchyma in vitro. We utilized a tissue engineering approach to develop a physiologically relevant model of glioma cell migration. This revealed that glioma cells display dramatic differences in migration when challenged by random versus aligned electrospun poly-ɛ-caprolactone nanofibers. Cells on aligned fibers migrated at an effective velocity of 4.2 ± 0.39 μm/h compared to 0.8 ± 0.08 μm/h on random fibers, closely matching in vivo models and prior observations of glioma spread in white versus gray matter. Cells on random fibers exhibited extension along multiple fiber axes that prevented net motion; aligned fibers promoted a fusiform morphology better suited to infiltration. Time-lapse microscopy revealed that the motion of individual cells was complex and was influenced by cell cycle and local topography. Glioma stem cell–containing neurospheres seeded on random fibers did not show cell detachment and retained their original shape; on aligned fibers, cells detached and migrated in the fiber direction over a distance sixfold greater than the perpendicular direction. This chemically and physically flexible model allows time-lapse analysis of glioma cell migration while recapitulating in vivo cell morphology, potentially allowing identification of physiological mediators and pharmacological inhibitors of invasion. PMID:19199562

The power of presence: the role of mindfulness at work for daily levels and change trajectories of psychological detachment and sleep quality.

PubMed

Hülsheger, Ute R; Lang, Jonas W B; Depenbrock, Franziska; Fehrmann, Carmen; Zijlstra, Fred R H; Alberts, Hugo J E M

2014-11-01

In this research, we examined the role of mindfulness for recovery from work using a daily diary design (N = 121; 5 days; 3 measurement occasions per day). The first goal of the study was to investigate the relationship of mindfulness with sleep quality and the mediating role of psychological detachment from a day-level perspective. A second goal was to extend the process perspective in recovery research beyond the day level and consider systematic change trajectories in recovery variables over the course of the work week and the role of mindfulness in these trajectories. Results regarding day-level relationships confirmed that mindfulness experienced during work was related to subsequent sleep quality, and this relationship was mediated by psychological detachment from work in the evening. Furthermore, an investigation of the role of mindfulness in recovery change trajectories supported the idea that psychological detachment trajectories increase over the work week for individuals low on mindfulness while there was no systematic mean-level change for individuals high on mindfulness. In contrast, sleep quality followed a linear increase from Monday to Friday for all individuals, irrespective of their levels of trait mindfulness. Practical and theoretical implications for the mindfulness and the recovery literature are discussed in conclusion. (PsycINFO Database Record (c) 2014 APA, all rights reserved).

A new scaling for divertor detachment

NASA Astrophysics Data System (ADS)

Goldston, R. J.; Reinke, M. L.; Schwartz, J. A.

2017-05-01

The ITER design, and future reactor designs, depend on divertor ‘detachment,’ whether partial, pronounced or complete, to limit heat flux to plasma-facing components and to limit surface erosion due to sputtering. It would be valuable to have a measure of the difficulty of achieving detachment as a function of machine parameters, such as input power, magnetic field, major radius, etc. Frequently the parallel heat flux, estimated typically as proportional to P sep/R or P sep B/R, is used as a proxy for this difficulty. Here we argue that impurity cooling is dependent on the upstream density, which itself must be limited by a Greenwald-like scaling. Taking this into account self-consistently, we find the impurity fraction required for detachment scales dominantly as power divided by poloidal magnetic field. The absence of any explicit scaling with machine size is concerning, as P sep surely must increase greatly for an economic fusion system, while increases in the poloidal field strength are limited by coil technology and plasma physics. This result should be challenged by comparison with 2D divertor codes and with measurements on existing experiments. Nonetheless, it suggests that higher magnetic field, stronger shaping, double-null operation, ‘advanced’ divertor configurations, as well as alternate means to handle heat flux such as metallic liquid and/or vapor targets merit greater attention.

A new scaling for divertor detachment

DOE PAGES

Goldston, R. J.; Reinke, M. L.; Schwartz, J. A.

2017-03-29

The ITER design, and future reactor designs, depend on divertor `detachment,'whether partial, pronounced or complete, to limit heat flux to plasma-facing components and to limit surface erosion due to sputtering. It would be valuable to have a measure of the difficulty of achieving detachment as a function of machine parameters, such as input power, magnetic field, major radius, etc. Frequently the parallel heat flux, estimated typically as proportional to P-sep/R or PsepB/R, is used as a proxy for this difficulty. Here we argue that impurity cooling is dependent on the upstream density, which itself must be limited by a Greenwald-likemore » scaling. Taking this into account self-consistently, we find the impurity fraction required for detachment scales dominantly as power divided by poloidal magnetic field. The absence of any explicit scaling with machine size is concerning, as P-sep surely must increase greatly for an economic fusion system, while increases in the poloidal field strength are limited by coil technology and plasma physics. This result should be challenged by comparison with 2D divertor codes and with measurements on existing experiments. Nonetheless, it suggests that higher magnetic field, stronger shaping, double-null operation, `advanced' divertor configurations, as well as alternate means to handle heat flux such as metallic liquid and/or vapor targets merit greater attention.« less

Creating technological boundaries to protect bedtime: examining work-home boundary management, psychological detachment and sleep.

PubMed

Barber, Larissa K; Jenkins, Jade S

2014-08-01

This study examined the mechanism by which information and communication technology (ICT) use at home for work purposes may affect sleep. In this investigation, data from 315 employees were used to examine the indirect effect of ICT use at home on sleep outcomes through psychological detachment, and how boundary creation may moderate this effect. Results revealed the indirect effect of increased work-home boundary crossing on sleep (quantity, quality and consistency) through psychological detachment occurred only among individuals with low boundaries around ICT use and not among those with high boundaries. These results suggest that creating boundaries around work-relevant ICT use while at home is beneficial to sleep as a recovery process through being able to psychologically disengage from work. Copyright © 2013 John Wiley & Sons, Ltd.

ELECTRON MICROSCOPIC STUDY OF CHANGES IN LIVER CELLS AFTER COMBINED TREATMENT WITH CARBON TETRACHLORIDE AND X RAYS (in Italian)

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

Longostrevi, G.P.; Lombardi, L.

Adult rats were treated with low doses of both CCl/sub 4/ and x rays, shown previously not to produce ultrastructural changes in liver parenchymal cells when given separately. The rats were exposed to a single, whole-body, 200- r dose of 160-kv x rays and then injected subcutaneously with 0.1 ml of a 30% solution of CCl/sub 4/ in peanut oil, corresponding to 0.15 ml CCl/sub 4/ per 100 g body wt. When the rats were sacrificed three days later, all showed pronounced changes in mitochondria and endoplasmic reticulum of liver cells, which were not seen following either individual treatment. Themore » mitochondria showed marked swelling, detachment of cristae, and accumulation of granular material in their interior. Endoplasmic reticulum underwent dilatation and detachment of normally adherent ribonucleoprotein granules. A different picture was observed following injection of toxic doses of CCl/sub 4/ (0.5 m1/100 g body wt). This produced marked disruption and ballooning of mitochondria and endoplasmic reticulum. The results indicate that subliminal doses of CCl/sub 4/ and x rays, presumably acting by different mechanisms, synergistically damage the liver cell, particularly its mitochondria. (BBB)« less

Morphodifferentiation of Gené's organ in engorged Amblyomma sculptum Berlese, 1888 female ticks (Acari: Ixodidae).

PubMed

Dos Santos, Marcelo Francisco; Maltauro Soares, Magna Aparecida; Lallo, Maria Anete; Barros-Battesti, Darci Moraes; de Lima-Netto, Solange; Spadacci-Morena, Diva Denelle

2018-03-01

The Gené's organ (GO) secretes a waxy substance on eggs that reduces water loss and has antimicrobial properties. The current study evaluated morphological and histochemical aspects of GO in Amblyomma sculptum from the period of post-feeding - when ticks detach from the host - to the stage just before oviposition. In this species, GO is composed of a corpus and two pairs of glands, namely, cranial and caudal. Glandular cells are joined laterally by a system of interdigitating membranes with junctional complexes. Histochemistry showed that lipid droplets became more evident as GO developed, while glycogen gradually disappeared, and proteins were detected only near the onset of oviposition. The ultrastructural results revealed a marked distension of the cuticle filled with an amorphous material. Glandular cells showed poor endoplasmatic reticulum, many mitochondria mainly in the basal cell poles and a very developed basal labyrinth. We concluded that the development of GO in A. sculptum ticks was continuous and progressive, and it started after detachment from the host. Additionally, the ultrastructure study suggests that gland cells have an important absorption ability and a low synthetic activity, which indicates that the majority of wax precursors are derived from haemolymph. Copyright © 2018 Elsevier GmbH. All rights reserved.

Selective killing of ovarian cancer cells through induction of apoptosis by nonequilibrium atmospheric pressure plasma

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

Iseki, Sachiko; Tanaka, Hiromasa; Kondo, Hiroki

2012-03-12

Two independent ovarian cancer cell lines and fibroblast controls were treated with nonequilibrium atmospheric pressure plasma (NEAPP). Most ovarian cancer cells were detached from the culture dish by continuous plasma treatment to a single spot on the dish. Next, the plasma source was applied over the whole dish using a robot arm. In vitro cell proliferation assays showed that plasma treatments significantly decreased proliferation rates of ovarian cancer cells compared to fibroblast cells. Flow cytometry and western blot analysis showed that plasma treatment of ovarian cancer cells induced apoptosis. NEAPP could be a promising tool for therapy for ovarian cancers.

Group choreography: mechanisms orchestrating the collective movement of border cells

PubMed Central

Montell, Denise J.; Yoon, Wan Hee; Starz-Gaiano, Michelle

2014-01-01

Cell movements are essential for animal development and homeostasis but also contribute to disease. Moving cells typically extend protrusions towards a chemoattractant, adhere to the substrate, contract and detach at the rear. It is less clear how cells that migrate in interconnected groups in vivo coordinate their behaviour and navigate through natural environments. The border cells of the Drosophila melanogaster ovary have emerged as an excellent model for the study of collective cell movement, aided by innovative genetic, live imaging, and photomanipulation techniques. Here we provide an overview of the molecular choreography of border cells and its more general implications. PMID:23000794

Modelling the detachment dependence on strike point location in the small angle slot divertor (SAS) with SOLPS

NASA Astrophysics Data System (ADS)

Casali, Livia; Covele, Brent; Guo, Houyang

2017-10-01

The new Small Angle Slot (SAS) divertor in DIII-D is characterized by a shallow-angle target enclosed by a slot structure about the strike point (SP). SOLPS modelling results of SAS have demonstrated divertor closure's utility in widening the range of acceptable densities for adequate heat handling. An extensive database of runs has been built to study the detachment dependence on SP location in SAS. Density scans show that lower Te at lower upstream density occur when the SP is at the critical location in the slot. The cooling front spreads across the entire target at higher densities, in agreement with experimental Langmuir probe measurements. A localized increase of the atomic and molecular density takes place near the SP, which reduces the target incident power density and facilitates detachment at lower upstream density. Systematic scans of variables such as power, transport, and viscosity have been carried out to assess the detachment sensitivity. Therein, a positive role of the viscosity is found. This work supported by DOE Contract Number DE-FC02-04ER54698.

Surface Charge and Hydrophobicity of Endospores of Bacillus anthracis and Related Species in Aqueous Solution

EPA Science Inventory

The surface properties of microorganisms play an important role in attachment and detachment in the environment. The change in surface charge can effect coagulation, disinfection, adhesion to surfaces, uptake of chemicals, and environmental transport. In aqueous solution, cell s...

Chalk-Ex: Transport of Optically Active Particles from the Surface Mixed Layer

DTIC Science & Technology

2002-09-30

and K. A. Kilpatrick. 1998. Scattering and attenuation properties of Emiliania huxleyi cells and their detached coccoliths. Limnol. Oceanogr. 43...coccolithophore Emiliania huxleyi under steady-state light-limited growth. Marine Ecology Progress Series. 142: 87-97. Bidigare, R. R. , M. Latasa, Z

Light Scattering by Marine Particles: Modeling with Non-spherical Shapes

DTIC Science & Technology

2008-01-01

Emiliania huxleyi, Limnology and Oceanography, 46, 1438−1454. Gordon, H.R., 2004, Inverse Radiative Transfer, Coccolith Backscattering, and Light Scattering...16430. Voss, K.J., W.M. Balch, and K.A. Kilpatrick, 1998, Scattering and attenuation properties of Emiliania huxleyi cells and their detached

Load and Pi control flux through the branched kinetic cycle of myosin V.

PubMed

Kad, Neil M; Trybus, Kathleen M; Warshaw, David M

2008-06-20

Myosin V is a processive actin-based motor protein that takes multiple 36-nm steps to deliver intracellular cargo to its destination. In the laser trap, applied load slows myosin V heavy meromyosin stepping and increases the probability of backsteps. In the presence of 40 mm phosphate (P(i)), both forward and backward steps become less load-dependent. From these data, we infer that P(i) release commits myosin V to undergo a highly load-dependent transition from a state in which ADP is bound to both heads and its lead head trapped in a pre-powerstroke conformation. Increasing the residence time in this state by applying load increases the probability of backstepping or detachment. The kinetics of detachment indicate that myosin V can detach from actin at two distinct points in the cycle, one of which is turned off by the presence of P(i). We propose a branched kinetic model to explain these data. Our model includes P(i) release prior to the most load-dependent step in the cycle, implying that P(i) release and load both act as checkpoints that control the flux through two parallel pathways.

pH dependent growth of poly( L-lysine)/poly( L-glutamic) acid multilayer films and their cell adhesion properties

NASA Astrophysics Data System (ADS)

Richert, Ludovic; Arntz, Youri; Schaaf, Pierre; Voegel, Jean-Claude; Picart, Catherine

2004-10-01

The short-term interaction of chondrosarcoma cells with (PGA/PLL) polyelectrolyte multilayers was investigated in a serum-containing medium for films built at different pHs and subsequently exposed to the culture medium. The buildup of the films and their stability was first investigated by means of optical waveguide lightmode spectroscopy, quartz crystal microbalance, streaming potential measurements and atomic force microscopy. While film growth is linear at all pHs, after a few layers have been deposited the growth is much larger for the films built at basic pH and even more pronounced for those built at acidic pH. However, these latter films remain stable in the culture medium only if they have been crosslinked prior to the ionic strength and pH jumps. The films built at acidic pH were found to swell in water by about 200% whereas those built at other pHs did not swell in a physiological buffer. For thin films (≈20 nm) built at pH = 7.4, the detachment forces were dependent on the outermost layer, the forces being significantly higher on PLL-ending films than on PGA-ending ones. In contrast, for the thick films built at pH = 4.4 and at pH = 10.4 (thickness of the order of few hundred of nanometers), the detachment forces were independent of the outermost layer of the film. The films built at pH = 10.4, which shrink in contact with salt containing solutions, were highly cell adhesive whereas those built at acidic pH were highly cell resistant. Protein adsorption and film roughness (as measured by AFM) could not explain these striking differences. The high adhesion observed on the film built at pH 10.4 may rather be related to the secondary structure of the film and to its relatively low swellability in water, whereas the cell resistance of the films built at pH 4.4 may be linked to their high swellability. Therefore, for the PGA/PLL films, the cell adhesion properties can be tuned depending on the deposition pH of the polyelectrolyte solutions. This study reveals the importance of the multilayer structure and architecture to control the detachment force of cells onto such films.

Inkjet-Print Micromagnet Array on Glass Slides for Immunomagnetic Enrichment of Circulating Tumor Cells

PubMed Central

Chen, Peng; Huang, Yu-Yen; Bhave, Gauri; Hoshino, Kazunori; Zhang, Xiaojing

2015-01-01

We report an inkjet-printed microscale magnetic structure that can be integrated on regular glass slides for the immunomagnetic screening of rare Circulating Tumor Cells (CTCs). CTCs detach from the primary tumor site, circulate with the bloodstream, and initiate the cancer metastasis process. Therefore, a liquid biopsy in the form of capturing and analyzing CTCs may provide key information for cancer prognosis and diagnosis. Inkjet printing technology provides a non-contact, layer-by-layer and mask-less approach to deposit defined magnetic patterns on an arbitrary substrate. Such thin film patterns, when placed in an external magnetic field, significantly enhance the attractive force in the near-field close to the CTCs to facilitate the separation. We demonstrated the efficacy of the inkjet-print micromagnet array integrated immunomagnetic assay in separating COLO205 (human colorectal cancer cell line) from whole blood samples. The micromagnets increased the capture efficiency by 26% compared with using plain glass slide as the substrate. PMID:26289942

Local 3D matrix microenvironment regulates cell migration through spatiotemporal dynamics of contractility-dependent adhesions

NASA Astrophysics Data System (ADS)

Doyle, Andrew D.; Carvajal, Nicole; Jin, Albert; Matsumoto, Kazue; Yamada, Kenneth M.

2015-11-01

The physical properties of two-dimensional (2D) extracellular matrices (ECMs) modulate cell adhesion dynamics and motility, but little is known about the roles of local microenvironmental differences in three-dimensional (3D) ECMs. Here we generate 3D collagen gels of varying matrix microarchitectures to characterize their regulation of 3D adhesion dynamics and cell migration. ECMs containing bundled fibrils demonstrate enhanced local adhesion-scale stiffness and increased adhesion stability through balanced ECM/adhesion coupling, whereas highly pliable reticular matrices promote adhesion retraction. 3D adhesion dynamics are locally regulated by ECM rigidity together with integrin/ECM association and myosin II contractility. Unlike 2D migration, abrogating contractility stalls 3D migration regardless of ECM pore size. We find force is not required for clustering of activated integrins on 3D native collagen fibrils. We propose that efficient 3D migration requires local balancing of contractility with ECM stiffness to stabilize adhesions, which facilitates the detachment of activated integrins from ECM fibrils.

A New Scaling for Divertor Detachment

NASA Astrophysics Data System (ADS)

Goldston, Robert

2017-10-01

The ITER design and future fusion power plant designs depend on divertor detachment, whether partial, pronounced or complete, both to limit heat flux to plasma-facing components and to limit surface erosion due to sputtering. Generally the parallel heat flux, estimated as proportional to Psep / R or Psep B / R , is used as a proxy for the difficulty of achieving detachment. Here we argue that the impurity cooling required for detachment is strongly dependent on the upstream separatrix density, which is limited by Greenwald scaling. Taking this into account self-consistently, along with the Heuristic Drift (HD) model for the SOL width, and using a Lengyel radiation model that includes non-coronal effects, we find that the relative impurity concentration, cz ≡nz /ne , required for detachment scales dominantly as cz Psep /Bp(nsep /nGW) 2 . The absence of any explicit favorable size scaling is concerning, as Psep must increase by an order of magnitude from present experiments to an economic fusion power system, while increases in the poloidal magnetic field strength are limited by magnet technology and MHD stability. This result should not be surprising, as it follows from the simplest scaling, Psep czne2VSOL , taking into account the Greenwald density limit and the HD SOL volume scaling. Reinke has combined a similar approach with the requirement to maintain H-mode, which sets a lower limit on Psep, and also arrives at an incentive for high field and disincentive for large size. These results should be challenged by comparison with 2D divertor codes and with measurements on existing experiments. In particular measurements are required for extrinsic divertor impurity concentration over a range of power and density conditions far from the regime where detachment can be achieved with deuterium puffing and intrinsic impurities alone. Nonetheless, these results suggest that higher magnetic field, stronger shaping, double-null operation, `advanced' divertor magnetic and baffle configurations, as well as lithium vapor targets merit greater attention. This work supported by the US DOE under contract DE-AC02-09CH11466.

LUT Reveals a New Mass-transferring Semi-detached Binary

NASA Astrophysics Data System (ADS)

Qian, S.-B.; Zhou, X.; Zhu, L.-Y.; Zejda, M.; Soonthornthum, B.; Zhao, E.-G.; Zhang, J.; Zhang, B.; Liao, W.-P.

2015-12-01

GQ Dra is a short-period eclipsing binary in a double stellar system that was discovered by Hipparcos. Complete light curves in the UV band were obtained with the Lunar-based Ultraviolet Telescope in 2014 November and December. Photometric solutions are determined using the W-D (Wilson and Devinney) method. It is discovered that GQ Dra is a classical Algol-type semi-detached binary where the secondary component is filling the critical Roche lobe. An analysis of all available times of minimum light suggests that the orbital period is increasing continuously at a rate of \\dot{P}=+3.48(+/- 0.23)× {10}-7 days yr-1. This could be explained by mass transfer from the secondary to the primary, which is in agreement with the semi-detached configuration with a lobe-filling secondary. By assuming a conservation of mass and angular momentum, the mass transfer rate is estimated as \\dot{m}=9.57(+/- 0.63)× {10}-8 {M}⊙ {{yr}}-1. All of these results reveal that GQ Dra is a mass-transferring semi-detached binary in a double system that was formed from an initially detached binary star. After the massive primary evolves to fill the critical Roche lobe, the mass transfer will be reversed and the binary will evolve into a contact configuration with two sub-giant or giant component stars.

Cohesiveness and hydrodynamic properties of young drinking water biofilms.

PubMed

Abe, Yumiko; Skali-Lami, Salaheddine; Block, Jean-Claude; Francius, Grégory

2012-03-15

Drinking water biofilms are complex microbial systems mainly composed of clusters of different size and age. Atomic force microscopy (AFM) measurements were performed on 4, 8 and 12 weeks old biofilms in order to quantify the mechanical detachment shear stress of the clusters, to estimate the biofilm entanglement rate ξ. This AFM approach showed that the removal of the clusters occurred generally for mechanical shear stress of about 100 kPa only for clusters volumes greater than 200 μm3. This value appears 1000 times higher than hydrodynamic shear stress technically available meaning that the cleaning of pipe surfaces by water flushing remains always incomplete. To predict hydrodynamic detachment of biofilm clusters, a theoretical model has been developed regarding the averaging of elastic and viscous stresses in the cluster and by including the entanglement rate ξ. The results highlighted a slight increase of the detachment shear stress with age and also the dependence between the posting of clusters and their volume. Indeed, the experimental values of ξ allow predicting biofilm hydrodynamic detachment with same order of magnitude than was what reported in the literature. The apparent discrepancy between the mechanical and the hydrodynamic detachment is mainly due to the fact that AFM mechanical experiments are related to the clusters local properties whereas hydrodynamic measurements reflected the global properties of the whole biofilm. Copyright © 2011 Elsevier Ltd. All rights reserved.

Psychological Detachment Mediating the Daily Relationship between Workload and Marital Satisfaction

PubMed Central

Germeys, Lynn; De Gieter, Sara

2017-01-01

Scholars already demonstrated that psychologically detaching from work after workhours can diminish or avoid the negative effects of job demands on employees' well-being. In this study, we examined a curvilinear relationship between workload and psychological detachment. Moreover, we investigated the moderating influence of an employee's work-home segmentation preference on the relation between detachment and marital satisfaction. In addition, we applied and extended the stressor-detachment model by examining detachment as a mediator of the relation between workload and marital satisfaction. A total of 136 employees participated in our daily diary survey study during 10 consecutive working days. The results of the Bayesian 2-level path analyses revealed a negative linear and curvilinear relationship between workload and psychological detachment on a daily basis. Daily detachment positively related to marital satisfaction, with one's preference to segment work from home reinforcing this relationship. Moreover, psychological detachment fully mediated the daily relationship between workload and marital satisfaction. Implications for practice and suggestions for future research are discussed. PMID:28101076

Bulk water phase and biofilm growth in drinking water at low nutrient conditions.

PubMed

Boe-Hansen, Rasmus; Albrechtsen, Hans-Jørgen; Arvin, Erik; Jørgensen, Claus

2002-11-01

In this study, the bacterial growth dynamics of a drinking water distribution system at low nutrient conditions was studied in order to determine bacterial growth rates by a range of methods, and to compare growth rates in the bulk water phase and the biofilm. A model distribution system was used to quantify the effect of retention times at hydraulic conditions similar to those in drinking water distribution networks. Water and pipe wall samples were taken and examined during the experiment. The pipes had been exposed to drinking water at approximately 13 degrees C, for at least 385 days to allow the formation of a mature quasi-stationary biofilm. At retention times of 12 h, total bacterial counts increased equivalent to a net bacterial growth rate of 0.048 day(-1). The bulk water phase bacteria exhibited a higher activity than the biofilm bacteria in terms of culturability, cell-specific ATP content, and cell-specific leucine incorporation rate. Bacteria in the bulk water phase incubated without the presence of biofilm exhibited a bacterial growth rate of 0.30 day(-1). The biofilm was radioactively labelled by the addition of 14C-benzoic acid. Subsequently, a biofilm detachment rate of 0.013 day(-1) was determined by measuring the release of 14C-labelled bacteria of the biofilm. For the quasi-stationary phase biofilm, the detachment rate was equivalent to the net growth rate. The growth rates determined in this study by different independent experimental approaches were comparable and within the range of values reported in the literature.

Rice Stomatal Closure Requires Guard Cell Plasma Membrane ATP-Binding Cassette Transporter RCN1/OsABCG5.

PubMed

Matsuda, Shuichi; Takano, Sho; Sato, Moeko; Furukawa, Kaoru; Nagasawa, Hidetaka; Yoshikawa, Shoko; Kasuga, Jun; Tokuji, Yoshihiko; Yazaki, Kazufumi; Nakazono, Mikio; Takamure, Itsuro; Kato, Kiyoaki

2016-03-07

Water stress is one of the major environmental stresses that affect agricultural production worldwide. Water loss from plants occurs primarily through stomatal pores. Here, we report that an Oryza sativa half-size ATP-binding cassette (ABC) subfamily G protein, RCN1/OsABCG5, is involved in stomatal closure mediated by phytohormone abscisic acid (ABA) accumulation in guard cells. We found that the GFP-RCN1/OsABCG5-fusion protein was localized at the plasma membrane in guard cells. The percentage of guard cell pairs containing both ABA and GFP-RCN1/OsABCG5 increased after exogenous ABA treatment, whereas they were co-localized in guard cell pairs regardless of whether exogenous ABA was applied. ABA application resulted in a smaller increase in the percentage of guard cell pairs containing ABA in rcn1 mutant (A684P) and RCN1-RNAi than in wild-type plants. Furthermore, polyethylene glycol (drought stress)-inducible ABA accumulation in guard cells did not occur in rcn1 mutants. Stomata closure mediated by exogenous ABA application was strongly reduced in rcn1 mutants. Finally, rcn1 mutant plants had more rapid water loss from detached leaves than the wild-type plants. These results indicate that in response to drought stress, RCN1/OsABCG5 is involved in accumulation of ABA in guard cells, which is indispensable for stomatal closure. Copyright © 2016 The Author. Published by Elsevier Inc. All rights reserved.

Femtosecond laser cutting of multiple thin corneal stromal lamellae for endothelial bioengineering.

PubMed

Bernard, Aurélien; He, Zhiguo; Forest, Fabien; Gauthier, Anne-Sophie; Peocʼh, Michel; Dumollard, Jean-Marc; Acquart, Sophie; Montard, Romain; Delbosc, Bernard; Gain, Philippe; Thuret, Gilles

2015-02-01

To assess the feasibility of cutting multiple thin stromal lamellae in human donor corneas using a commercial femtosecond laser (FSL) to provide cell carriers for future endothelial graft bioengineering. Eight edematous organ-cultured corneas not suitable for grafting for endothelial reasons were mounted on a Ziemer anterior chamber and cut with a Z6 FSL with 6 successive parallel cuts, from depth to surface. Target thickness of each lamella ranged from 100 to 150 μm depending on initial corneal thickness. Thickness was measured using anterior segment optical coherence tomography before and after cutting on mounted corneas, and on each stromal lamella after detachment. Scanning electron microscopy observation was performed on 4 lamellae and histological cross sections on 1 cornea before detachment. A median of 5 (minimum 3, maximum 7) lamellae was obtained per cornea. All lamellae still attached were the most posterior ones, suggesting that FSL was less efficient because of light scattering by edematous stroma. Cut precision and postdetachment swelling were correlated with anterior-posterior position within the cornea. Median lamella thickness was 127 μm (56-222 μm) before detachment and 196 μm (80-304 μm) after detachment. Surface state was consistent with previously reported FSL lamellar cuts during Descemet stripping automated endothelial keratoplasty. Up to 7 thin lamellae can be cut in stored corneas with an FSL. This method, once optimized primarily by using deswelled, more transparent corneas, could prove effective for recycling unsuitable donor corneas in corneal bioengineering processes.

Psychological Detachment in the Relationship between Job Stressors and Strain

PubMed Central

Safstrom, My; Hartig, Terry

2013-01-01

We investigated the mediating versus moderating role of psychological detachment in the relationship between job stressors and psychological strain. Our sample consisted of 173 university students invested in challenging programs of advanced professional studies, who could find it difficult to detach from work. Hierarchical regression analyses of cross-sectional survey data affirmed the role of psychological detachment as a mediator in the relationship between job demands and perceived stress. Detachment also mediated the relationship between job demands and satisfaction with life, although the association disappeared when controlling for negative affectivity. Detachment did not mediate relationships between job demands and cognitive failures. Psychological detachment did not moderate any of the investigated relationships. The study contributes to a view of psychological detachment as less subject to individual differences than to the imposition of stressors in the given context. PMID:25379246

Bi-Directional Reflectance Measurements of the Ocean Bottom

DTIC Science & Technology

2001-09-30

waters”, 1998, Limnol. and Ocean., 43: 1198 - 1206. K J. Voss, W. M. Balch, K. A. Kilpatrick, "Scattering and attenuation properties of Emiliania ... huxleyi cells and their detached coccoliths", 1998, Limnol. and Ocean., 43: 870-876. J. S. Bartlett, K. J. Voss, S. Sathyendranath, and A. Vodacek

Molecular Alterations that Reduce Cortical Containment of Tubulin Microtentacles and their Impact on Breast Tumor Metastasis

DTIC Science & Technology

2010-04-01

This technique provides a reliable method of determining cellular plasticity and metastatic potential [5, 6]. These studies are nearing completion and...directed chemotherapies could influence CTCs and high- lights the need to investigate these effects. Current methods for detecting CTCs in metastatic...understanding of how these compounds influence detached and circulating tumor cells. Materials and methods Cell culture MCF-10A human MECs were

Clustering of brain tumor cells: a first step for understanding tumor recurrence

NASA Astrophysics Data System (ADS)

Khain, Evgeniy; Nowicki, M. O.; Chiocca, E. A.; Lawler, S. E.; Schneider-Mizell, C. M.; Sander, L. M.

2012-02-01

Glioblastoma tumors are highly invasive; therefore the overall prognosis of patients remains poor, despite major improvements in treatment techniques. Cancer cells detach from the inner tumor core and actively migrate away [1]; eventually these invasive cells might form clusters, which can develop to recurrent tumors. In vitro experiments in collagen gel [1] followed the clustering dynamics of different glioma cell lines. Based on the experimental data, we formulated a stochastic model for cell dynamics, which identified two mechanisms of clustering. First, there is a critical value of the strength of adhesion; above the threshold, large clusters grow from a homogeneous suspension of cells; below it, the system remains homogeneous, similarly to the ordinary phase separation. Second, when cells form a cluster, there is evidence that their proliferation rate increases. We confirmed the theoretical predictions in a separate cell migration experiment on a substrate and found that both mechanisms are crucial for cluster formation and growth [2]. In addition to their medical importance, these phenomena present exciting examples of pattern formation and collective cell behavior in intrinsically non-equilibrium systems [3]. [4pt] [1] A. M. Stein et al, Biophys. J., 92, 356 (2007). [0pt] [2] E. Khain et al, EPL 88, 28006 (2009). [0pt] [3] E. Khain et al, Phys. Rev. E. 83, 031920 (2011).

Ultra-fast microwave-assisted hydrothermal synthesis of long vertically aligned ZnO nanowires for dye-sensitized solar cell application.

PubMed

Mahpeykar, S M; Koohsorkhi, J; Ghafoori-Fard, H

2012-04-27

Long vertically aligned ZnO nanowire arrays were synthesized using an ultra-fast microwave-assisted hydrothermal process. Using this method, we were able to grow ZnO nanowire arrays at an average growth rate as high as 200 nm min(-1) for maximum microwave power level. This method does not suffer from the growth stoppage problem at long growth times that, according to our investigations, a normal microwave-assisted hydrothermal method suffers from. Longitudinal growth of the nanowire arrays was investigated as a function of microwave power level and growth time using cross-sectional FESEM images of the grown arrays. Effect of seed layer on the alignment of nanowires was also studied. X-ray diffraction analysis confirmed c-axis orientation and single-phase wurtzite structure of the nanowires. J-V curves of the fabricated ZnO nanowire-based mercurochrome-sensitized solar cells indicated that the short-circuit current density is increased with increasing the length of the nanowire array. According to the UV-vis spectra of the dyes detached from the cells, these increments were mainly attributed to the enlarged internal surface area and therefore dye loading enhancement in the lengthened nanowire arrays.

Force-velocity relationship of single actin filament interacting with immobilised myosin measured by electromagnetic technique.

PubMed

Holohan, S-J P; Marston, S B

2005-06-01

The effect of applying an external load to actin filaments moving in the in vitro motility assay is studied. Bead-tailed actin filaments were made by polymerising actin onto 2.8 microm diameter Dynabeads conjugated with gelsolin-G actin. These were introduced into a motility cell coated with 100 microg/ml rabbit fast skeletal myosin in the presence of ATP and 0.5% methylcellulose. The motility cell was inserted between the pole-pieces of an electromagnet and the fluorescent beads and filaments were observed. The force-current relationship of the electromagnet was determined from the velocity of free beads in viscous solution and Stokes' equation. The magnet produced up to 6 pN force on the Dynabeads at 1 A. Many bead-tailed actin filaments stuck to the surface, but the beads that did move moved at the same speed as unloaded f-actin in the same cell. Bead-tailed filaments slowed down under an increasing magnetic load, eventually stalled and then slid backward under increasing load before detaching from the surface. Single-filament force-velocity curves were constructed and a stalling force of about 0.6 pN/mm of actin filament estimated.

The detached haze layer in Titan's mesosphere: The formation process

NASA Astrophysics Data System (ADS)

Lavvas, P.; Yelle, R. V.; Vuitton, V.

2008-09-01

Cassini observations made by the Imaging Science Subsystem (ISS) [1] and by the UltraViolet Imaging Spectrometer (UVIS) [2,3] have revealed the presence of a detached haze layer in Titan's mesosphere at an altitude of 520 km. Analysis of the observed optical properties presented in the accompanying talk [5], suggests that the average size of particles in the detached layer is of ~40 nm, with an imaginary index k < 0.3 at 187.5 nm and a number density of ˜30 particles cm-3, while calculations of the sedimentation velocity of the haze particles coupled with the derived number density imply a mass flux of 1.9-3.2 × 10-14 g cm-2 s-1. This is approximately equal to the mass flux required to explain the main haze layer and suggests that the main haze layer in Titan's stratosphere is formed primarily by sedimentation and coagulation of particles in the detached layer [5,6]. The HASI data clearly show that the haze is coincident with a temperature maximum. This rules out condensation as the source of the detached haze. We have also considered a more complicated scenario in which the detached layer is caused by an increase in the density of condensation nuclei near 520 km. This is motivated by the fact that silicate micrometeorites ablate near 500 km [7,8]. Recondensation of the refractory vapor creates `smoke' particles that could serve as condensation nuclei. Combination of Pioneer measurements along with theoretical estimations for the particles velocity distribution, suggest a mass flux of ~10-17 g cm-2 s-1 at Saturn's region [9], while measurements from the Cassini Dust Analyser (CDA) suggest a similar magnitude at Titan's location [10]. These fluxes are ~3 orders of magnitude smaller than the lower limit of the estimated mass flux out of the detached haze layer, so meteorite ablation can not be the direct cause of the aerosol layer. However, if the ablated meteoritic material reforms 1 nm particles, the implied number flux would be 2.4 × 103 particles cm-2 s-1, which is of the right order of magnitude to explain the detached layer. This hypothesis requires that additional material condense on the meteoritic smoke particles. Unfortunately, the main photochemical products on Titan (HCN, C2H2, C2H6, etc.) do not condense at the temperature and pressure in the detached layer. The saturation mixing ratio for species present in Titan's mesosphere are shown in Fig. 1. The vapour pressure of each species is calculated assuming the HASI vertical temperature profile [4]. The species that come closest to condensing are H2O and C6N2. There is some water vapor present from ablation of icy micrometeorites in Titan's atmosphere, but the mole fraction corresponding to saturation vapor pressure of water at 520 km is 1.6 × 10-2, many orders of magnitude larger than expected [11]. Similarly, the mole fraction of C6N2 at 520 km is expected to be much smaller than the saturation value of 5 × 10-6 [12]. Hence, the growth of particles through condensation cannot explain the detached haze layer. Advection processes in the atmosphere have been related to the formation of the Voyager detached haze layer [13]. In this picture, meridional winds transport the haze particles polewards, constraining them at a specific altitude region, before depositing them at the pole, while the upwelling part of the circulation transports large particles from the main haze layer upwards, enhancing in this way the opacity of the detached haze layer. Yet, the meridional winds are estimated to be υ~3 cm s-1 in the region of the stratospheric zonal jet near 0.1 mbar based on CIRS measurements [14]. Assuming this value to hold in the region of the detached haze layer implies a horizontal motion characteristic time of H~R/v = 108 s. A 40 nm particle has a settling velocity of υ S~1 cm s-1 at 500 km, and the characteristic time to fall 20 km is only 2 × 106 s implying that the particles fall out of the region before they are transported to the pole. Thus, the dynamical explanation for the Voyager detached haze fails by more than a factor of 50 for the detached layer at 520 km. The inefficiency of the above processes to provide enough mass flux to generate the detached layer along with the large mass flux retrieved by the observations [5,6] suggest that haze is formed in Titan's thermosphere by high-energy radical and ion chemistry. In this case the presence of the detached layer could be an optical illusion effect due to the increase of particles' size, along with the decrease of their settling velocity with decreasing altitude. In order to investigate this effect we used a microphysical model [15], which has been extended to describe both spherical and fractal particle growth [16]. We assume a gaussian production profile centered at 800 km and with a column mass production of 4.5 × 10-14 g cm-2 s-1. The particles grow as spheres or fractals depending on the fractal dimension, Df, (Df=2 for fractals and 3 for spheres). Using only spherical particle growth provides a good fit to the extinction observed by UVIS at 187.5 nm [3] above the detached haze layer, while the modeled extinction drops faster than the observed extinction at altitudes below the detached layer (Fig. 2). By setting the transition altitude of spherical to fractal growth at 510 km, the generated fractal particles provide an excellent fit to the observed extinction profile below the detached layer suggesting that the particles present there are indeed of fractal structure. At the same time, the transition from spheres to fractals aggregates, generates a well pronounced minimum in the total simulated extinction, which provides a very good fit to the observed detached haze layer extinction profile [3]. In addition, the calculated mass flux at 520 km is 3.1 × 10-14 g cm-2 s-1, which is within the limits defined by the optical analysis of the detached layer [5,6]. Finally, the calculated monomer size of the particles forming the fractal aggregates is 57 nm, is good agreement with the 50 nm size retrieved by the DISR measurements [17]. Our simulation of Titan's haze particle evolution suggests that the presence of the detached haze layer is due to the transition in the growth of particles from spherical to fractal structure. Further investigation of the processes defining the growth of the particles is required in order to understand why the transition takes place at this region and how the particles produced at higher altitudes are defining the vertical haze opacity in Titan's atmosphere.

Connexin-43 upregulation in micrometastases and tumor vasculature and its role in tumor cell attachment to pulmonary endothelium

PubMed Central

Elzarrad, M Khair; Haroon, Abu; Willecke, Klaus; Dobrowolski, Radoslaw; Gillespie, Mark N; Al-Mehdi, Abu-Bakr

2008-01-01

Background The modulation of gap junctional communication between tumor cells and between tumor and vascular endothelial cells during tumorigenesis and metastasis is complex. The notion of a role for loss of gap junctional intercellular communication in tumorigenesis and metastasis has been controversial. While some of the stages of tumorigenesis and metastasis, such as uncontrolled cell division and cellular detachment, would necessitate the loss of intercellular junctions, other stages, such as intravasation, endothelial attachment, and vascularization, likely require increased cell-cell contact. We hypothesized that, in this multi-stage scheme, connexin-43 is centrally involved as a cell adhesion molecule mediating metastatic tumor attachment to the pulmonary endothelium. Methods Tumor cell attachment to pulmonary vasculature, tumor growth, and connexin-43 expression was studied in metastatic lung tumor sections obtained after tail-vein injection into nude mice of syngeneic breast cancer cell lines, overexpressing wild type connexin-43 or dominant-negatively mutated connexin-43 proteins. High-resolution immunofluorescence microscopy and Western blot analysis was performed using a connexin-43 monoclonal antibody. Calcein Orange Red AM dye transfer by fluorescence imaging was used to evaluate the gap junction function. Results Adhesion of breast cancer cells to the pulmonary endothelium increased with cancer cells overexpressing connexin-43 and markedly decreased with cells expressing dominant-negative connexin-43. Upregulation of connexin-43 was observed in tumor cell-endothelial cell contact areas in vitro and in vivo, and in areas of intratumor blood vessels and in micrometastatic foci. Conclusion Connexin-43 facilitates metastatic 'homing' by increasing adhesion of cancer cells to the lung endothelial cells. The marked upregulation of connexin-43 in tumor cell-endothelial cell contact areas, whether in preexisting 'homing' vessels or in newly formed tumor vessels, suggests that connexin-43 can serve as a potential marker of micrometastases and tumor vasculature and that it may play a role in the early incorporation of endothelial cells into small tumors as seeds for vasculogenesis. PMID:18647409

Connexin-43 upregulation in micrometastases and tumor vasculature and its role in tumor cell attachment to pulmonary endothelium.

PubMed

Elzarrad, M Khair; Haroon, Abu; Willecke, Klaus; Dobrowolski, Radoslaw; Gillespie, Mark N; Al-Mehdi, Abu-Bakr

2008-07-22

The modulation of gap junctional communication between tumor cells and between tumor and vascular endothelial cells during tumorigenesis and metastasis is complex. The notion of a role for loss of gap junctional intercellular communication in tumorigenesis and metastasis has been controversial. While some of the stages of tumorigenesis and metastasis, such as uncontrolled cell division and cellular detachment, would necessitate the loss of intercellular junctions, other stages, such as intravasation, endothelial attachment, and vascularization, likely require increased cell-cell contact. We hypothesized that, in this multi-stage scheme, connexin-43 is centrally involved as a cell adhesion molecule mediating metastatic tumor attachment to the pulmonary endothelium. Tumor cell attachment to pulmonary vasculature, tumor growth, and connexin-43 expression was studied in metastatic lung tumor sections obtained after tail-vein injection into nude mice of syngeneic breast cancer cell lines, overexpressing wild type connexin-43 or dominant-negatively mutated connexin-43 proteins. High-resolution immunofluorescence microscopy and Western blot analysis was performed using a connexin-43 monoclonal antibody. Calcein Orange Red AM dye transfer by fluorescence imaging was used to evaluate the gap junction function. Adhesion of breast cancer cells to the pulmonary endothelium increased with cancer cells overexpressing connexin-43 and markedly decreased with cells expressing dominant-negative connexin-43. Upregulation of connexin-43 was observed in tumor cell-endothelial cell contact areas in vitro and in vivo, and in areas of intratumor blood vessels and in micrometastatic foci. Connexin-43 facilitates metastatic 'homing' by increasing adhesion of cancer cells to the lung endothelial cells. The marked upregulation of connexin-43 in tumor cell-endothelial cell contact areas, whether in preexisting 'homing' vessels or in newly formed tumor vessels, suggests that connexin-43 can serve as a potential marker of micrometastases and tumor vasculature and that it may play a role in the early incorporation of endothelial cells into small tumors as seeds for vasculogenesis.

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

Takahashi, Eri, E-mail: eritakahashi@fc.kuh.kumamoto-u.ac.jp; Fukushima, Ayako; Haga, Akira

In rhegmatogenous retinal detachment (RRD), scattered RPE cells from the basement membrane into the vitreous cavity undergo an epithelial mesenchymal transition (EMT) and form the intraocular fibrous membrane in response to vitreous fluid. We investigated whether exposure to vitreous samples was associated with EMT-associated signals and mesenchymal characters. Human vitreous samples were collected from patients with RRD, epiretinal membrane (ERM), or macular hole (MH). We evaluated the effects of vitreous on ARPE-19 cells in suspension cultures using poly 2-hydroxyethyl methacrylate-coated dishes and three-dimensional (3D) Matrigel cultures. We found that exposure to vitreous samples did not induce morphological changes or accelerate woundmore » closure in monolayers. Several samples showed increased phosphorylation of Smad2 and nuclear translocation of nuclear factor-κB. Mechanical stress triggered an elevation of phosphorylation levels in Smad2. In addition, exposure to vitreous fluid increased the phosphorylation of p38 mitogen-activated protein kinase in cell suspension cultures after mechanical stress. Moreover, ARPE-19 cells showed a stellate invasive phenotype in 3D Matrigel cultures with vitreous samples. In this study, we demonstrated that mechanical stress and vitreous were associated with EMT-associated signals and invasive phenotypes in 3D cultures but not in monolayers. These results have important implications for the role of vitreous humor in the induction of EMT and intraocular fibrosis.« less

Reduction of Defects in Germanium-Silicon

NASA Technical Reports Server (NTRS)

2003-01-01

Crystals grown without contact with a container have far superior quality to otherwise similar crystals grown in direct contact with a container. In addition to float-zone processing, detached- Bridgman growth is a promising tool to improve crystal quality, without the limitations of float zoning or the defects introduced by normal Bridgman growth. Goals of this project include the development of the detached Bridgman process to be reproducible and well understood and to quantitatively compare the defect and impurity levels in crystals grown by these three methods. Germanium (Ge) and germanium-silicon (Ge-Si) alloys are being used. At MSFC, we are responsible for the detached Bridgman experiments intended to differentiate among proposed mechanisms of detachment, and to confirm or refine our understanding of detachment. Because the contact angle is critical to determining the conditions for detachment, the sessile drop method was used to measure the contact angles as a function of temperature and composition for a large number of substrates made of potential ampoule materials. Growth experiments have used pyrolytic boron nitride (pBN) and fused silica ampoules with the majority of the detached results occurring predictably in the pBN. Etch pit density (EPD) measurements of normal and detached Bridgman-grown Ge samples show a two order of magnitude improvement in the detached-grown samples. The nature and extent of detachment is determined by using profilometry in conjunction with optical and electron microscopy. The stability of detachment has been analyzed, and an empirical model for the conditions necessary to achieve sufficient stability to maintain detached growth for extended periods has been developed. We have investigated the effects on detachment of ampoule material, pressure difference above and below the melt, and Si concentration; samples that are nearly completely detached can be grown repeatedly in pBN. Current work is concentrated on developing a method to make in situ pressure measurements in the growth ampoules.

Happy, healthy, and productive: the role of detachment from work during nonwork time.

PubMed

Fritz, Charlotte; Yankelevich, Maya; Zarubin, Anna; Barger, Patricia

2010-09-01

Mentally distancing oneself from work during nonwork time can help restore resources lost because of work demands. In this study, we examined possible outcomes of such psychological detachment from work, specifically well-being and job performance. Although employees may need to mentally detach from work to restore their well-being, high levels of detachment may require a longer time to get back into "working mode," which may be negatively associated with job performance. Our results indicate that higher levels of self-reported detachment were associated with higher levels of significant other-reported life satisfaction as well as lower levels of emotional exhaustion. In addition, we found curvilinear relationships between psychological detachment and coworker reported job performance (task performance and proactive behavior). Thus, although high psychological detachment may enhance employee well-being, it seems that medium levels of detachment are most beneficial for job performance. Copyright 2010 APA, all rights reserved

Quaternary low-angle slip on detachment faults in Death Valley, California

USGS Publications Warehouse

Hayman, N.W.; Knott, J.R.; Cowan, D.S.; Nemser, E.; Sarna-Wojcicki, A. M.

2003-01-01

Detachment faults on the west flank of the Black Mountains (Nevada and California) dip 29??-36?? and cut subhorizontal layers of the 0.77 Ma Bishop ash. Steeply dipping normal faults confined to the hanging walls of the detachments offset layers of the 0.64 Ma Lava Creek B tephra and the base of 0.12-0.18 Ma Lake Manly gravel. These faults sole into and do not cut the low-angle detachments. Therefore the detachments accrued any measurable slip across the kinematically linked hanging-wall faults. An analysis of the orientations of hundreds of the hanging-wall faults shows that extension occurred at modest slip rates (<1 mm/yr) under a steep to vertically oriented maximum principal stress. The Black Mountain detachments are appropriately described as the basal detachments of near-critical Coulomb wedges. We infer that the formation of late Pleistocene and Holocene range-front fault scarps accompanied seismogenic slip on the detachments.

Fine Tuning Cell Migration by a Disintegrin and Metalloproteinases

PubMed Central

Theodorou, K.

2017-01-01

Cell migration is an instrumental process involved in organ development, tissue homeostasis, and various physiological processes and also in numerous pathologies. Both basic cell migration and migration towards chemotactic stimulus consist of changes in cell polarity and cytoskeletal rearrangement, cell detachment from, invasion through, and reattachment to their neighboring cells, and numerous interactions with the extracellular matrix. The different steps of immune cell, tissue cell, or cancer cell migration are tightly coordinated in time and place by growth factors, cytokines/chemokines, adhesion molecules, and receptors for these ligands. This review describes how a disintegrin and metalloproteinases interfere with several steps of cell migration, either by proteolytic cleavage of such molecules or by functions independent of proteolytic activity. PMID:28260841

A Study of Parameters Affecting Fibroblast Morphology in Response to an Applied Mechanical Force

NASA Technical Reports Server (NTRS)

Grymes, Rosalind A.; Sawyer, Christine

1994-01-01

A precisely controlled stretch/relaxation regimen (20% elongation at 6.6 cycles/min) was applied to normal human fetal, neonatal and aged dermal fibroblasts cultured on flexible membranes. Culture conditions included poly (NH2) or collagen type I coated substrate membranes; control cultures were grown on the same pliable material in the absence of applied stretch. Direct observation and immunofluorescence analyses revealed a progressive change in cell body orientation limited to the stretched dermal fibroblast cultures. Monolayers gradually (over 4 days) acquired a symmetric, radial distribution equivalent to the biaxial array of the applied force. At high seeding density, alignment was inhibited in the fetal cell cultures. This cell strain required collagen type I coating for optimal attachment to the flexible membrane, preferring growth in three-dimensional cell 'balls' on the poly(NH2) coated substrate. Neonatal cells also required the collagen type I coating, but both neonatal and aged dermal fibroblasts aligned efficiently at all seeding densities examined. The randomly oriented neonatal cells on the unstretched control membranes spontaneously detached at confluence, as a single cell sheet. Their aligned counterparts did not detach until the applied stretch stimulus was removed. Low concentrations of cytochalasin D (62.5 ng/ml) disrupted the stretch-related alignment response. Rhodamine phalloidin staining visualized fewer actin stress fibers in stretched, aligned cells than in controls. Both intercellular interactions and cytoskeletal integrity mediate the response to mechanical strain. Normal rabbit corneal stroma fibroblasts (NRC) were also analyzed, and failed to orient under these conditions. This cell type may require a different regimen, or a longer time period, to demonstrate alignment behavior. Supported by NASA Space Biology RTOP 199-40-22 and the NASA-ARC Director's Discretionary Fund.

Delineation of the vitreous and posterior hyaloid using bromophenol blue.

PubMed

Haritoglou, Christos; Strauss, Rupert; Priglinger, Siegfried G; Kreutzer, Thomas; Kampik, Anselm

2008-02-01

To describe visualization of the vitreous and the posterior hyaloid membrane using bromophenol blue during vitrectomy for macular hole and retinal detachment. Six patients with macular holes and four with retinal detachments were included in the study. Before and after surgery, complete clinical examination, including funduscopy and measurements of best-corrected visual acuity and intraocular pressure, was performed. Additional functional tests, such as fluorescein angiography, optical coherence tomography (Stratus OCT; Carl Zeiss Meditec, Jena, Germany, Germany), Goldmann perimetry, and multifocal electroretinography as well as photography of the posterior pole, were performed for macular hole patients. Bromophenol blue was used in concentrations of 0.2%. During macular hole surgery, the dye was injected into the air-filled globe, while during surgery for retinal detachment, the globe was partially filled with perfluorocarbon before dye injection after induction of a posterior vitreous detachment to stain the vitreous peripherally. Bromophenol blue provided sufficient staining of the attached posterior hyaloid membrane and vitreous remnants in the periphery. This was especially helpful for patients in whom a posterior vitreous detachment could not be induced mechanically by suction using the vitrectomy probe alone, as seen in three of six interventions for a macular hole in this series. In addition, staining of the vitreous or vitreous remnants in the periphery and at the vitreous base was seen in all patients and helped to completely remove the vitreous in a controlled fashion. After macular hole surgery, increase of visual acuity from 20/100 (mean) to 20/40 was seen during follow-up up to 6 months. In one case, the hole persisted and required a second operation. Finally, closure of the hole was achieved in all patients. After retinal detachment surgery, reattachment was achieved in all cases. No dye-related adverse events were seen during follow-up as shown by the functional tests (visual acuity measurement, electroretinography, and perimetry) applied. Delineation of the vitreous and the posterior hyaloid using bromophenol blue staining greatly facilitates vitreoretinal procedures. Bromophenol blue appeared to be a very helpful and safe tool to visualize the posterior hyaloid membrane in macular hole surgery and assured its complete separation from the retinal surface. The dye also helped to remove vitreous at the vitreous base during retinal detachment surgery. Therefore, bromophenol blue appears as a very good alternative to triamcinolone, which has been used for this purpose, because the dye has no pharmacological properties and no side effects are likely to occur such as cataract formation and increase in intraocular pressure. Further studies including larger numbers of patients are mandatory.

Impact of grain size and rock composition on simulated rock weathering

NASA Astrophysics Data System (ADS)

Israeli, Yoni; Emmanuel, Simon

2018-05-01

Both chemical and mechanical processes act together to control the weathering rate of rocks. In rocks with micrometer size grains, enhanced dissolution at grain boundaries has been observed to cause the mechanical detachment of particles. However, it remains unclear how important this effect is in rocks with larger grains, and how the overall weathering rate is influenced by the proportion of high- and low-reactivity mineral phases. Here, we use a numerical model to assess the effect of grain size on chemical weathering and chemo-mechanical grain detachment. Our model shows that as grain size increases, the weathering rate initially decreases; however, beyond a critical size no significant decrease in the rate is observed. This transition occurs when the density of reactive boundaries is less than ˜ 20 % of the entire domain. In addition, we examined the weathering rates of rocks containing different proportions of high- and low-reactivity minerals. We found that as the proportion of low-reactivity minerals increases, the weathering rate decreases nonlinearly. These simulations indicate that for all compositions, grain detachment contributes more than 36 % to the overall weathering rate, with a maximum of ˜ 50 % when high- and low-reactivity minerals are equally abundant in the rock. This occurs because selective dissolution of the high-reactivity minerals creates large clusters of low-reactivity minerals, which then become detached. Our results demonstrate that the balance between chemical and mechanical processes can create complex and nonlinear relationships between the weathering rate and lithology.

Methionine Deprivation Induces a Targetable Vulnerability in Triple-Negative Breast Cancer Cells by Enhancing TRAIL Receptor-2 Expression.

PubMed

Strekalova, Elena; Malin, Dmitry; Good, David M; Cryns, Vincent L

2015-06-15

Many neoplasms are vulnerable to methionine deficiency by mechanisms that are poorly understood. Because gene profiling studies have revealed that methionine depletion increases TNF-related apoptosis-inducing ligand receptor-2 (TRAIL-R2) mRNA, we postulated that methionine stress sensitizes breast cancer cells to proapoptotic TRAIL-R2 agonists. Human triple (ER/PR/HER2)-negative breast carcinoma cell lines were cultured in control or methionine-free media. The effects of methionine depletion on TRAIL receptor expression and sensitivity to chemotherapy or a humanized agonistic TRAIL-R2 monoclonal antibody (lexatumumab) were determined. The melanoma-associated antigen MAGED2 was silenced to delineate its functional role in sensitizing TNBC cells to methionine stress. An orthotopic TNBC model was utilized to evaluate the effects of dietary methionine deficiency, lexatumumab, or the combination. Methionine depletion sensitized TNBC cells to lexatumumab-induced caspase activation and apoptosis by increasing TRAIL-R2 mRNA and cell surface expression. MCF-10A cells transformed by oncogenic H-Ras, but not untransformed cells, and matrix-detached TNBC cells were highly sensitive to the combination of lexatumumab and methionine depletion. Proteomics analyses revealed that MAGED2, which has been reported to reduce TRAIL-R2 expression, was suppressed by methionine stress. Silencing MAGED2 recapitulated features of methionine deprivation, including enhanced mRNA and cell surface expression of TRAIL receptors and increased sensitivity to TRAIL receptor agonists. Dietary methionine deprivation enhanced the antitumor effects of lexatumumab in an orthotopic metastatic TNBC model. Methionine depletion exposes a targetable defect in TNBC cells by increasing TRAIL-R2 expression. Our findings provide the foundation for a clinical trial combining dietary methionine restriction and TRAIL-R2 agonists. Clin Cancer Res; 21(12); 2780-91. ©2015 AACR. ©2015 American Association for Cancer Research.

Renal myofibroblasts contract collagen I matrix lattices in vitro.

PubMed

Kelynack, K J; Hewitson, T D; Pedagogos, E; Nicholls, K M; Becker, G J

1999-01-01

Myofibroblasts, cells with both fibroblastic and smooth muscle cell features, have been implicated in renal scarring. In addition to synthetic properties, contractile features and integrin expression may allow myofibroblasts to rearrange and contract interstitial collagenous proteins. Myofibroblasts from normal rat kidneys were grown in cell-populated collagen lattices to measure cell generated contraction. Following detachment of cell populated collagen lattices, myofibroblasts progressively contracted collagen lattices, reducing lattice diameter by 42% at 24 h. Alignment of myofibroblasts, rearrangement of fibrils and beta(1) integrin expression were observed within lattices. We postulate that interstitial myofibroblasts contribute to renal scarring through manipulation of collagenous proteins. Copyright 1999 S. Karger AG, Basel

Capture and release of cells using a temperature-responsive surface that immobilizes an antibody through DNA duplex formation.

PubMed

Kimura, Tsuyoshi; Nakamura, Naoko; Umeda, Kanji; Hashimoto, Yoshihide; Kishida, Akio

We synthesized a temperature-responsive surface that immobilized an antibody via DNA duplex formation for selective capture and release of target cells. Polyethylene films were modified by grafting poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAAm-co-AAc)), which were prepared at various ratios of NIPAAm/AAc. The increased hydrophilicity of P(NIPAAm-co-PAA) film with decreased temperature was confirmed by water contact angle measurement. Single strand DNA (20mer) was chemically immobilized on the surface and then antibody (anti-mouse CD45, mCD45) modified with the complementary single strand DNA was immobilized on the surface through DNA duplex formation. The mCD45 antibody immobilization was confirmed by immunostaining. HeLa cells (mCD45 negative) and mouse bone marrow (BM) cells (mCD45 positive) were adhered on the surfaces at 37 °C. Although HeLa cells were detached by 4 °C incubation, BM cells were still adhered on the surface and then the adhered cells were released by DNase treatment. From these results, it was suggested that cells could be selectively captured and collected by using a film having surface that immobilizes an antibody via DNA duplex formation.

Treatment of in vitro enterohemorrhagic Escherichia coli infection using phage and probiotics.

PubMed

Dini, C; Bolla, P A; de Urraza, P J

2016-07-01

To assay the combination of phage and probiotics against EHEC in vitro on infected Hep-2 cells. Phage and probiotics treatments on EHEC O157:H7-infected Hep-2 cells were assayed individually or combined. The effect of freeze-drying on phage and probiotic antimicrobial activity was also studied. While treatment with phage alone increased cell detachment caused by EHEC infection, the treatments with MM alone or in combination with phage proved to effectively diminish cell damage caused by EHEC infection. Combined treatment showed a decrease in apoptotic cell count of 57·3% and a reduction in EHEC adhesion to cell monolayer of 1·2 log CFU. The simultaneous use of phage and probiotics showed no antagonistic effect, and freeze-drying did not affect their antipathogenic activity. The combination of phage and probiotics has great potential for reducing the number of pathogens adhered to epithelial cells during EHEC O157:H7 infection and attenuating the cytotoxic effect derived from it. Further in vivo assays are needed for assessing the actual effectiveness of the treatment. This study presents a freeze-dried formulation of phage and probiotics capable of controlling EHEC infections and reducing epithelial cell damage in vitro. © 2016 The Society for Applied Microbiology.

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

Yang, Lu; Hanson, David E

Here we present the results on the study of surface properties of {beta}-HMX crystal utilizing molecular simulations. The surface polarity of three principal crystal surfaces are investigated by measuring the water contact angles. The calculated contact angles agree excellently with the values measured by experiment and show that the surface polarity of three crystal surfaces are different. The free energies and forces of detaching an Estane chain with and without nitroplasticizer from the three principal crystal surfaces were calculated using umbrella sampling technique. We find that the detaching free energy/force increases with the increasing HMX surface polarity. In addition, ourmore » results also show that nitroplasticizer plays an important role in the adhesion forces between Estane and HMX surfaces.« less

StarD13 is a tumor suppressor in breast cancer that regulates cell motility and invasion

PubMed Central

HANNA, SAMER; KHALIL, BASSEM; NASRALLAH, ANITA; SAYKALI, BECHARA A.; SOBH, RANIA; NASSER, SELIM; EL-SIBAI, MIRVAT

2014-01-01

Breast cancer is one of the most commonly diagnosed cancers in women around the world. In general, the more aggressive the tumor, the more rapidly it grows and the more likely it metastasizes. Members of the Rho subfamily of small GTP-binding proteins (GTPases) play a central role in breast cancer cell motility and metastasis. The switch between active GTP-bound and inactive GDP-bound state is regulated by guanine nucleotide exchange factors (GEFs), GTPase-activating proteins (GAPs) and guanine-nucleotide dissociation inhibitors (GDIs). We studied the role of StarD13, a recently identified Rho-GAP that specifically inhibits the function of RhoA and Cdc42. We aimed to investigate its role in breast cancer proliferation and metastasis. The levels of expression of this Rho-GAP in tumor tissues of different grades were assayed using immunohistochemistry. We observed that, while the level of StarD13 expression decreases in cancer tissues compared to normal tissues, it increases as the grade of the tumor increased. This was consistent with the fact that although StarD13 was indeed a tumor suppressor in our breast cancer cells, as seen by its effect on cell proliferation, it was needed for cancer cell motility. In fact, StarD13 knockdown resulted in an inhibition of cell motility and cells were not able to detach their tail and move forward. Our study describes, for the first time, a tumor suppressor that plays a positive role in cancer motility. PMID:24627003

Detachment of secondary dendrite arm in a directionally solidified Sn-Ni peritectic alloy under deceleration growth condition

PubMed Central

Peng, Peng; Li, Xinzhong; Li, Jiangong; Su, Yanqing; Guo, Jingjie; Fu, Hengzhi

2016-01-01

In order to better understand the detachment mechanism of secondary dendrite arm during peritectic solidification, the detachment of secondary dendrite arm from the primary dendrite arms in directionally solidified Sn-36at.%Ni peritectic alloys is investigated at different deceleration rates. Extensive detachment of secondary dendrite arms from primary stem is observed below peritectic reaction temperature TP. And an analytical model is established to characterize the detachment process in terms of the secondary dendrite arm spacing λ2, the root radius of detached arms and the specific surface area (SV) of dendrites. It is found that the detachment mechanism is caused by not only curvature difference between the tips and roots of secondary branches, but also that between the thicker secondary branches and the thinner ones. Besides, this detachment process is significantly accelerated by the temperature gradient zone melting (TGZM) effect during peritectic solidification. It is demonstrated that the reaction constant (f) which is used to characterize the kinetics of peritectic reaction is crucial for the determination of the detachment process. The value of f not only changes with growth rate but also with solidification time at a given deceleration rate. In conclusion, these findings help the better understanding of the detachment mechanism. PMID:27270334

An Assessment of Molten Metal Detachment Hazards During Electron Beam Welding in Space

NASA Technical Reports Server (NTRS)

Fragomeni, James M.; Nunes, Arthur C., Jr.

1998-01-01

The safety issue has been raised with regards to potential molten metal detachments from the weld pool and cold filler wire during electron beam welding in space. This investigation was undertaken to evaluate if molten metal could detach and come in contact with astronauts and burn through the fabric of the astronauts' Extravehicular Mobility Unit (EMU) during electron beam welding in space. Molten metal detachments from either the weld/cut substrate or weld wire could present harm to a astronaut if the detachment was to burn through the fabric of the EMU. Theoretical models were developed to predict the possibility and size of the molten metal detachment hazards during the electron beam welding exercises at Low Earth Orbit (LEO). The primary molten metal detachment concerns were those cases of molten metal separation from the metal surface due to metal cutting, weld pool splashing, entrainment and release of molten metal due to filler wire snap-out from the weld puddle, and molten metal accumulation and release from the end of the weld wire. Some possible ways of obtaining molten metal drop detachments would include an impulse force, or bump, to the weld sample, cut surface, or filler wire. Theoretical models were developed for these detachment concerns from principles of impact and kinetic energies, surface tension, drop geometry, surface energies, and particle dynamics. The surface tension represents the force opposing the liquid metal drop from detaching whereas the weight of the liquid metal droplet represents a force that is tending to detach the molten metal drop. Theoretical calculations have indicated that only a small amount of energy is required to detach a liquid metal drop; however, much of the energy of an impact is absorbed in the sample or weld plate before it reaches the metal drop on the cut edge or surface. The tendency for detachment is directly proportional to the weld pool radius and metal density and inversely proportional to the surface tension of the liquid metal. For a detachment the initial kinetic energy of the weld pool with respect to the plate has to exceed the energy to form the extra surface required for the detachment of the pool. The difficulty is in transferring the energy from the point of impact through the plate and sample to the cut edge. It is likely that not all of the kinetic energy is available for detaching the pool; some may be sequestered in weld pool oscillations. The coefficient of restitution for the collision will be lower than one if irreversible deformation, for example plastic flow deformation, takes place during the collision. Thus determining the amount of energy from an impact that actually reaches the molten metal droplet is critical. Various molten metal detachment scenarios were tested experimentally in an enclosed vacuum chamber using the Ukrainian Universal Hand Tool, an electron beam welder designed for space welding. The experimental testing was performed in a 4 ft. X 4 ft. vacuum chamber at Marshall Space Flight Center, evacuated to vacuum levels of at least 50 microTorr, and also some welding garment material was utilized to observe the effect of the molten metal detachments on the material. A "carillon" apparatus consisting of four pendulum hammer strikers, each weighing approximately 3.65 lbs, raised to predetermined specific heights was used to apply an impact force to the weld sample/plate during electron beam welding and cutting exercises. The strikers were released by switching on an electric motor to rotate a pin holding wires retaining the strikers at desired heights. The specimens were suspended so as to be free to respond to the blows with a sudden velocity increment. The specimens were mounted on a hinged plate for minimizing effective mass with the option to fasten it down so as to raise its effective mass closer to that anticipated for an actual space welding scenario. Measurements were made of the impact energy and the horizontal fling distances of the detached metal drops. It was not particularly easy to generate the detachments for this experiment. This document presents the details of the theoretical modeling effort and a summary of the experimental effort to measure molten metal drop detachments from terrestrial electron beam welding in the enclosed vacuum chamber. The results of the experimental effort have shown that molten metal detachments can occur from the sample/weld plate only if a sufficiently large impact force is applied to the weld plate. A "weld pool detachment parameter" was determined to indicate whether detachment would occur. Detachment can be either full or partial (dripping), Partial detachment means that the weld pool detached from one side of the liquid-solid boundary so as to leave a hole at the puddle site but remained attached over part of the liquid-solid boundary and dripped down the plate with no fully detached material detected. Full detachment, however, does not necessarily mean that the whole pool fully detached; in some cases only a smaller portion of the pool detached, the remainder dripping down the plate. The weld pool detachment parameter according to theory and according to the empirical data allows a determination of whether full detachments might occur. Theoretical calculations indicated titanium alloy would be the most difficult from which to detach molten metal droplets followed by stainless steel and then by aluminum. The experimental results were for the most part consistent with the theoretical analysis and predictions. The above theory is applicable to other situations as desired for assessing the potential for molten metal detachments.

Eye-Directed Overpressure Airwave-Induced Trauma Causes Lasting Damage to the Anterior and Posterior Globe: A Model for Testing Cell-Based Therapies

PubMed Central

Bricker-Anthony, Courtney; Hines-Beard, Jessica

2016-01-01

Abstract Purpose: Characterization of the response of the Balb/c mouse to an eye-directed overpressure airwave, with the hypothesis that this mouse strain and model is useful for testing potential therapeutics for the treatment of traumatic eye injury. Methods: The left eyes of adult Balb/c mice were exposed to an eye-directed overpressure airwave. Intraocular pressure (IOP) was measured and eyes were inspected for gross pathology changes. Optical coherence tomography and histology were used to examine the structural integrity of the retina and optic nerve. Immunohistochemistry, in vivo molecular fluorophores, and a multiplex enzyme-linked immunosorbent assay were utilized to identify changes in cell death, neuroinflammation, and oxidative stress. Results: This model induced a transient increase in IOP, corneal injuries, infrequent large retinal detachments, retinal pigment epithelium (RPE) vacuolization, glial reactivity, and retinal cell death. Both the corneal damage and RPE vacuolization persisted with time. Optic nerve degeneration occurred as early as 7 days postinjury and persisted out to 60 days. Retinal cell death, increased levels of reactive oxygen species, and neuroinflammation were detected at 7 days postinjury. Conclusions: The injury profile of the Balb/c mouse is consistent with commonly observed pathologies in blast-exposed patients. The damage is throughout the eye and persistent, making this mouse model useful for testing cell-based therapies. PMID:26982447

[Two cases of Vogt-Koyanagi-Harada disease presenting shallow anterior chamber].

PubMed

Takemoto, Daisuke; Ijiri, Shigeyuki; Shimizu, Michiharu; Higashide, Tomomi; Sugiyama, Kazuhisa

2015-05-01

We report two cases of Vogt-Koyanagi-Harada disease (VKH) in which shallow anterior chambers were improved after steroid pulse therapy. The patients were women aged 65 and 72. They had headaches, decreased visual acuity and shallow anterior chamber in both eyes. There was no inflammation in the anterior chamber. Ultrasound biomicroscopy (UBM) showed ciliary edema, ciliochoroidal detachment, and angle closure. One case showed high intraocular pressure (IOP), and a diagnosis of acute primary angle closure was made. Although cataract surgery was performed in the left eye, postoperative optical coherence tomography (OCT) revealed serous retinal detachment in both eyes. The shallow anterior chamber and UBM findings were improved and serous retinal detachment disappeared after steroid pulse therapy in both cases. VKH may cause shallow anterior chamber and angle closure. The inflammatory changes of VKH in the anterior segment, i. e. ciliary edema and ciliochoroidal detachment, may exacerbate the shallow anterior chambers and narrow angles and result in an acute increase in IOP in eyes with short axial length. VKH associated with shallow anterior chamber may be misdiagnosed as acute primary angle closure. For differential diagnosis, examinations of the ocular fundus including OCT are useful.

Early simultaneous fundus autofluorescence and optical coherence tomography features after pars plana vitrectomy for primary rhegmatogenous retinal detachment.

PubMed

Dell'Omo, Roberto; Mura, Marco; Lesnik Oberstein, Sarit Y; Bijl, Heico; Tan, H Stevie

2012-04-01

To describe fundus autofluorescence and optical coherence tomography (OCT) features of the macula after pars plana vitrectomy for rhegmatogenous retinal detachment. Thirty-three eyes of 33 consecutive patients with repaired rhegmatogenous retinal detachment with or without the involvement of the macula were prospectively investigated with simultaneous fundus autofluorescence and OCT imaging using the Spectralis HRA+OCT (Heidelberg Engineering, Heidelberg, Germany) within a few weeks after the operation. Fundus autofluorescence imaging of the macula showed lines of increased and decreased autofluorescence in 19 cases (57.6%). On OCT, these lines corresponded to the following abnormalities: outer retinal folds, inner retinal folds, and skip reflectivity abnormalities of the photoreceptor inner segment/outer segment band. Other OCT findings, not related to abnormal lines on fundus autofluorescence, consisted of disruption of photoreceptor inner segment/outer segment band and collection of intraretinal or subretinal fluid. The presence of outer retinal folds significantly related to metamorphopsia but did not relate to poor postoperative visual acuity. Partial-thickness retinal folds occur commonly after vitrectomy for rhegmatogenous retinal detachment repair and may represent an important anatomical substrate for postoperative metamorphopsia. Fundus autofluorescence and OCT are both sensitive techniques for the detection of these abnormalities.

Aberrant Apoptotic Response of Colorectal Cancer Cells to Novel Nucleoside Analogues.

PubMed

Harmse, Leonie; Dahan-Farkas, Nurit; Panayides, Jenny-Lee; van Otterlo, Willem; Penny, Clement

2015-01-01

Despite the increased understanding of colorectal cancer and the introduction of targeted drug therapy, the metastatic phase of the disease remains refractory to treatment. Since the deregulation of normal apoptosis contributes to the pathogenesis of colorectal cancer, novel nucleoside analogues were synthesized here and evaluated for their ability to induce apoptosis and cause cell death in two colorectal adeno-carcinoma cell lines, Caco-2 and HT-29. Three novel nucleoside analogues assessed here showed cytotoxic activity, as measured by the MTT assay against both cell lines: the IC50 values ranged between 3 and 37 μM, with Caco-2 cells being more sensitive than HT-29 cells. Compared to camptothecin, the positive control, the nucleoside analogues were significantly less toxic to normal unstimulated leukocytes (p>0.05). Moreover, the nucleosides were able to induce apoptosis as measured by an increase in caspase 8 and caspase 3 activity above that of the control. This was additionally supported by data derived from Annexin V-FITC assays. Despite marginal changes to the mitochondrial membrane potential, all three nucleosides caused a significant increase in cytosolic cytochrome c (p>0.05), with a corresponding decrease in mitochondrial cytochrome c. Morphological analysis of both cell lines showed the rapid appearance of vacuoles following exposure to two of the nucleosides, while a third caused cellular detachment, delayed cytoplasmic vacuolisation and nuclear abnormalities. Preliminary investigations, using the autophagic indicator monodansylcadaverine and chloroquine as positive control, showed that two of the nucleosides induced the formation of autophagic vacuoles. In summary, the novel nucleoside analogues showed selective cytotoxicity towards both cancer cell lines and are effective initiators of an unusual apoptotic response, demonstrating their potential to serve as structural scaffolds for more potent analogues.

The microenvironment induces collective migration in SDHB-silenced mouse pheochromocytoma spheroids.

PubMed

D'Antongiovanni, Vanessa; Martinelli, Serena; Richter, Susan; Canu, Letizia; Guasti, Daniele; Mello, Tommaso; Romagnoli, Paolo; Pacak, Karel; Eisenhofer, Graeme; Mannelli, Massimo; Rapizzi, Elena

2017-10-01

Pheochromocytomas (Pheos) and paragangliomas (PGLs) are neuroendocrine tumors. Approximately 30-40% of Pheos/PGLs are due to germline mutations in one of the susceptibility genes, including those encoding the succinate dehydrogenase subunits A-D ( SDHA-D ). Up to 2/3 of patients affected by SDHB mutated Pheo/PGL develop metastatic disease with no successful cure at present. Here, for the first time, we evaluated the effects of SDHB silencing in a three dimension (3D) culture using spheroids of a mouse Pheo cell line silenced or not (wild type/wt/control) for the SDHB subunit. We investigated the role of the microenvironment on spheroid growth and migration/invasion by co-culturing SDHB -silenced or wt spheroids with primary cancer-activated fibroblasts (CAFs). When spheroids were co-cultured with fibroblasts, SDHB -silenced cells showed a significant increase in matrigel invasion as demonstrated by the computation of the migratory areas ( P  < 0.001). Moreover, cells detaching from the SDHB -silenced spheroids moved collectively, unlike the cells of wt spheroids that moved individually. Additionally, SDHB- silenced spheroids developed long filamentous formations along which clusters of cells migrated far away from the spheroid, whereas these structures were not present in wt spheroids. We found that lactate, largely secreted by CAFs, plays a specific role in promoting migration only of SDHB -silenced cells. In this study, we demonstrated that SDHB silencing per se increases tumor cell migration/invasion and that microenvironment, as represented by CAFs, plays a pivotal role in enhancing collective migration/invasion in Pheo SDHB -silenced tumor cells, suggesting their role in increasing the tumor metastasizing potential. © 2017 Society for Endocrinology.

The RhoA Activator GEF-H1/Lfc Is a Transforming Growth Factor-β Target Gene and Effector That Regulates α-Smooth Muscle Actin Expression and Cell Migration

PubMed Central

Tsapara, Anna; Luthert, Phillip; Greenwood, John; Hill, Caroline S.

2010-01-01

Maintenance of the epithelial phenotype is crucial for tissue homeostasis. In the retina, dedifferentiation and loss of integrity of the retinal pigment epithelium (RPE) leads to retinal dysfunction and fibrosis. Transforming growth factor (TGF)-β critically contributes to RPE dedifferentiation and induces various responses, including increased Rho signaling, up-regulation of α-smooth muscle actin (SMA), and cell migration and dedifferentiation. Cellular TGF-β responses are stimulated by different signal transduction pathways: some are Smad dependent and others Smad independent. Alterations in Rho signaling are crucial to both types of TGF-β signaling, but how TGF-β-stimulates Rho signaling is poorly understood. Here, we show that primary RPE cells up-regulated GEF-H1 in response to TGF-β. GEF-H1 was the only detectable Rho exchange factor increased by TGF-β1 in a genome-wide expression analysis. GEF-H1 induction was Smad4-dependant and led to Rho activation. GEF-H1 inhibition counteracted α-SMA up-regulation and cell migration. In patients with retinal detachments and fibrosis, migratory RPE cells exhibited increased GEF-H1 expression, indicating that induction occurs in diseased RPE in vivo. Our data indicate that GEF-H1 is a target and functional effector of TGF-β by orchestrating Rho signaling to regulate gene expression and cell migration, suggesting that it represents a new marker and possible therapeutic target for degenerative and fibrotic diseases. PMID:20089843

Failed CTL/NK cell killing and cytokine hypersecretion are directly linked through prolonged synapse time

PubMed Central

Rudd-Schmidt, Jesse A.; Lopez, Jamie A.; Ramsbottom, Kelly M.; Mannering, Stuart I.; Andrews, Daniel M.; Voskoboinik, Ilia

2015-01-01

Failure of cytotoxic T lymphocytes (CTLs) or natural killer (NK) cells to kill target cells by perforin (Prf)/granzyme (Gzm)-induced apoptosis causes severe immune dysregulation. In familial hemophagocytic lymphohistiocytosis, Prf-deficient infants suffer a fatal “cytokine storm” resulting from macrophage overactivation, but the link to failed target cell death is not understood. We show that prolonged target cell survival greatly amplifies the quanta of inflammatory cytokines secreted by CTLs/NK cells and that interferon-γ (IFN-γ) directly invokes the activation and secondary overproduction of proinflammatory IL-6 from naive macrophages. Furthermore, using live cell microscopy to visualize hundreds of synapses formed between wild-type, Prf-null, or GzmA/B-null CTLs/NK cells and their targets in real time, we show that hypersecretion of IL-2, TNF, IFN-γ, and various chemokines is linked to failed disengagement of Prf- or Gzm-deficient lymphocytes from their targets, with mean synapse time increased fivefold, from ∼8 to >40 min. Surprisingly, the signal for detachment arose from the dying target cell and was caspase dependent, as delaying target cell death with various forms of caspase blockade also prevented their disengagement from fully competent CTLs/NK cells and caused cytokine hypersecretion. Our findings provide the cellular mechanism through which failed killing by lymphocytes causes systemic inflammation involving recruitment and activation of myeloid cells. PMID:25732304

``Sheddable'' PEG-lipid to balance the contradiction of PEGylation between long circulation and poor uptake

NASA Astrophysics Data System (ADS)

Zhao, Caiyan; Deng, Hongzhang; Xu, Jing; Li, Shuyi; Zhong, Lin; Shao, Leihou; Wu, Yan; Liang, Xing-Jie

2016-05-01

PEGylated lipids confer longer systemic circulation and tumor accumulation via the enhanced permeability and retention (EPR) effect. However, PEGylation inhibits cellular uptake and subsequent endosomal escape. In order to balance the contradiction between the advantages of long circulation and the disadvantages of poor uptake of PEGylated lipids, we prepared a ``sheddable'' PEG-lipid micelle system based on the conjugation of PEG and phosphatidyl ethanolamine (DSPE) with a pH sensitive benzoic imine bond. In a physiological environment, the PEG-protected micelles were not readily taken up by the reticuloendothelial system (RES) and could be successfully delivered to tumor tissue by the EPR effect. In a tumor acidic microenvironment, the PEG chains detached from the surfaces of the micelles while the degree of linker cleavage could not cause a significant particle size change, which facilitated the carrier binding to tumor cells and improved the cellular uptake. Subsequently, the ``sheddable'' PEG-lipid micelles easily internalized into cells and the increased acidity in the lysosomes further promoted drug release. Thus, this ``sheddable'' PEG-lipid nanocarrier could be a good candidate for effective intracellular drug delivery in cancer chemotherapy.PEGylated lipids confer longer systemic circulation and tumor accumulation via the enhanced permeability and retention (EPR) effect. However, PEGylation inhibits cellular uptake and subsequent endosomal escape. In order to balance the contradiction between the advantages of long circulation and the disadvantages of poor uptake of PEGylated lipids, we prepared a ``sheddable'' PEG-lipid micelle system based on the conjugation of PEG and phosphatidyl ethanolamine (DSPE) with a pH sensitive benzoic imine bond. In a physiological environment, the PEG-protected micelles were not readily taken up by the reticuloendothelial system (RES) and could be successfully delivered to tumor tissue by the EPR effect. In a tumor acidic microenvironment, the PEG chains detached from the surfaces of the micelles while the degree of linker cleavage could not cause a significant particle size change, which facilitated the carrier binding to tumor cells and improved the cellular uptake. Subsequently, the ``sheddable'' PEG-lipid micelles easily internalized into cells and the increased acidity in the lysosomes further promoted drug release. Thus, this ``sheddable'' PEG-lipid nanocarrier could be a good candidate for effective intracellular drug delivery in cancer chemotherapy. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr02174c

Aluminium oxide nanoparticles induced morphological changes, cytotoxicity and oxidative stress in Chinook salmon (CHSE-214) cells.

PubMed

Srikanth, Koigoora; Mahajan, Amit; Pereira, Eduarda; Duarte, Armando Costa; Venkateswara Rao, Janapala

2015-10-01

Aluminium oxide nanoparticles (Al2 O3 NPs) are increasingly used in diverse applications that has raised concern about their safety. Recent studies suggested that Al2 O3 NPs induced oxidative stress may be the cause of toxicity in algae, Ceriodaphnia dubia, Caenorhabditis elegans and Danio rerio. However, there is paucity on the toxicity of Al2 O3 NPs on fish cell lines. The current study was aimed to investigate Al2 O3 NPs induced cytotoxicity, oxidative stress and morphological abnormality of Chinnok salmon cells (CHSE-214). A dose-dependent decline in cell viability was observed in CHSE-214 cells exposed to Al2 O3 NPs. Oxidative stress induced by Al2 O3 NPs in CHSE-214 cells has resulted in the significant reduction of superoxide dismutase, catalase and glutathione in a dose-dependent manner. However, a significant increase in glutathione sulfo-transferase and lipid peroxidation was observed in CHSE-214 cells exposed to Al2 O3 NPs in a dose-dependent manner. Significant morphological changes in CHSE-214 cells were observed when exposed to Al2 O3 NPs at 6, 12 and 24 h. The cells started to detach and appear spherical at 6 h followed by loss of cellular contents resulting in the shrinking of the cells. At 24 h, the cells started to disintegrate and resulted in cell death. Our data demonstrate that Al2 O3 NPs induce cytotoxicity and oxidative stress in a dose-dependent manner in CHSE-214 cells. Thus, our current work may serve as a base-line study for future evaluation of toxicity studies using CHSE-214 cells. Copyright © 2015 John Wiley & Sons, Ltd.

A simple 2D biofilm model yields a variety of morphological features.

PubMed

Hermanowicz, S W

2001-01-01

A two-dimensional biofilm model was developed based on the concept of cellular automata. Three simple, generic processes were included in the model: cell growth, internal and external mass transport and cell detachment (erosion). The model generated a diverse range of biofilm morphologies (from dense layers to open, mushroom-like forms) similar to those observed in real biofilm systems. Bulk nutrient concentration and external mass transfer resistance had a large influence on the biofilm structure.

Relative Contributions of Various Cellular Mechanisms to Loss of Algae during Cnidarian Bleaching.

PubMed

Bieri, Tamaki; Onishi, Masayuki; Xiang, Tingting; Grossman, Arthur R; Pringle, John R

2016-01-01

When exposed to stress such as high seawater temperature, corals and other cnidarians can bleach due to loss of symbiotic algae from the host tissue and/or loss of pigments from the algae. Although the environmental conditions that trigger bleaching are reasonably well known, its cellular and molecular mechanisms are not well understood. Previous studies have reported the occurrence of at least four different cellular mechanisms for the loss of symbiotic algae from the host tissue: in situ degradation of algae, exocytic release of algae from the host, detachment of host cells containing algae, and death of host cells containing algae. The relative contributions of these several mechanisms to bleaching remain unclear, and it is also not known whether these relative contributions change in animals subjected to different types and/or durations of stresses. In this study, we used a clonal population of the small sea anemone Aiptasia, exposed individuals to various precisely controlled stress conditions, and quantitatively assessed the several possible bleaching mechanisms in parallel. Under all stress conditions tested, except for acute cold shock at 4°C, expulsion of intact algae from the host cells appeared to be by far the predominant mechanism of bleaching. During acute cold shock, in situ degradation of algae and host-cell detachment also became quantitatively significant, and the algae released under these conditions appeared to be severely damaged.

Relative Contributions of Various Cellular Mechanisms to Loss of Algae during Cnidarian Bleaching

PubMed Central

Bieri, Tamaki; Onishi, Masayuki; Xiang, Tingting; Grossman, Arthur R.; Pringle, John R

2016-01-01

When exposed to stress such as high seawater temperature, corals and other cnidarians can bleach due to loss of symbiotic algae from the host tissue and/or loss of pigments from the algae. Although the environmental conditions that trigger bleaching are reasonably well known, its cellular and molecular mechanisms are not well understood. Previous studies have reported the occurrence of at least four different cellular mechanisms for the loss of symbiotic algae from the host tissue: in situ degradation of algae, exocytic release of algae from the host, detachment of host cells containing algae, and death of host cells containing algae. The relative contributions of these several mechanisms to bleaching remain unclear, and it is also not known whether these relative contributions change in animals subjected to different types and/or durations of stresses. In this study, we used a clonal population of the small sea anemone Aiptasia, exposed individuals to various precisely controlled stress conditions, and quantitatively assessed the several possible bleaching mechanisms in parallel. Under all stress conditions tested, except for acute cold shock at 4°C, expulsion of intact algae from the host cells appeared to be by far the predominant mechanism of bleaching. During acute cold shock, in situ degradation of algae and host-cell detachment also became quantitatively significant, and the algae released under these conditions appeared to be severely damaged. PMID:27119147

Safety of vitrectomy for floaters.

PubMed

Tan, H Stevie; Mura, Marco; Lesnik Oberstein, Sarit Y; Bijl, Heico M

2011-06-01

To assess the risks of vitrectomy for the removal of primary and secondary vitreous opacities. Retrospective, nonrandomized, interventional case series. We reviewed the results of 116 consecutive cases of vitrectomy for vitreous floaters. Eighty-six cases were primary and 30 cases were secondary floaters. Main outcome measures were the incidence of iatrogenic retinal breaks and postoperative rhegmatogenous retinal detachments. We found iatrogenic retinal breaks in 16.4% of operations. There was no statistically significant difference in risk between cases of primary and secondary floaters. Intraoperative posterior vitreous detachment induction was found to increase significantly the risk of breaks. Retinal detachment occurred in 3 cases (2.5%), all after operations for primary floaters. One case of complicated retinal detachment ended with a low visual acuity of hand movements. Cataract occurred in 50% of phakic cases. Transient postoperative hypotony was found after 5.2% of our operations, and transient postoperative high intraocular pressure was encountered in 7.8%. An intraoperative choroidal hemorrhage occurred in 1 case, which resolved spontaneously. The mean visual acuity improved from 0.20 to 0.13 logarithm of the minimal angle of resolution units. The risk profile of vitrectomy for floaters is comparable with that of vitrectomy for other elective indications. Retinal breaks are a common finding during surgery and treatment of these breaks is crucial for the prevention of postoperative retinal detachment. Patients considering surgery for floaters should be informed specifically about the risks involved. Copyright © 2011 Elsevier Inc. All rights reserved.

Divertor detachment

NASA Astrophysics Data System (ADS)

Krasheninnikov, Sergei

2015-11-01

The heat exhaust is one of the main conceptual issues of magnetic fusion reactor. In a standard operational regime the large heat flux onto divertor target reaches unacceptable level in any foreseeable reactor design. However, about two decades ago so-called ``detached divertor'' regimes were found. They are characterized by reduced power and plasma flux on divertor targets and look as a promising solution for heat exhaust in future reactors. In particular, it is envisioned that ITER will operate in a partly detached divertor regime. However, even though divertor detachment was studied extensively for two decades, still there are some issues requiring a new look. Among them is the compatibility of detached divertor regime with a good core confinement. For example, ELMy H-mode exhibits a very good core confinement, but large ELMs can ``burn through'' detached divertor and release large amounts of energy on the targets. In addition, detached divertor regimes can be subject to thermal instabilities resulting in the MARFE formation, which, potentially, can cause disruption of the discharge. Finally, often inner and outer divertors detach at different plasma conditions, which can lead to core confinement degradation. Here we discuss basic physics of divertor detachment including different mechanisms of power and momentum loss (ionization, impurity and hydrogen radiation loss, ion-neutral collisions, recombination, and their synergistic effects) and evaluate the roles of different plasma processes in the reduction of the plasma flux; detachment stability; and an impact of ELMs on detachment. We also evaluate an impact of different magnetic and divertor geometries on detachment onset, stability, in- out- asymmetry, and tolerance to the ELMs. Supported by the U.S. Department of Energy Office of Science, Office of Fusion Energy Sciences under Award Number DE-DE-FG02-04ER54739 at UCSD.

Evaluation of hyaloid-retinal relationship during triamcinolone-assisted vitrectomy for primary rhegmatogenous retinal detachment.

PubMed

Sundar, Dheepak; Takkar, Brijesh; Venkatesh, Pradeep; Chawla, Rohan; Temkar, Shreyas; Azad, Shorya Vardhan; Vohra, Rajpal

2018-03-01

To determine hyaloid-retinal relationship in primary rhegmatogenous retinal detachment during vitreous surgery. This is a prospective, interventional study of patients (n = 72) undergoing triamcinolone-assisted 25G vitreous surgery for primary rhegmatogenous retinal detachment. Hyaloid-retinal relationship was noted intraoperatively to identify regions and patterns of firm attachment and was classified into subgroups. Analysis was done to determine association between hyaloid-retinal relationship patterns and preoperative findings: posterior vitreous detachment, proliferative vitreoretinopathy, type of retinal tear, the presence of peripheral degenerations, and postoperative outcomes. Three patterns of hyaloid-retinal relationship were identified: type1 (complete absence of posterior vitreous detachment (21%)), type 2 (incomplete posterior vitreous detachment (47%)) and type 3 (complete posterior vitreous detachment (32%)). Posterior vitreous detachment in some form was present in 84% of the cases with retinal tears as the causative break but none of the cases with retinal holes (p < 0.001). None of the cases with vitreoretinal degeneration had complete posterior vitreous detachment (p = 0.001). 69% of proliferative vitreoretinopathy-C cases had type 1 hyaloid-retinal relationship as compared to 11% cases with no proliferative vitreoretinopathy (p < 0.001). Proliferative vitreoretinopathy-related anatomical failure was seen in 7.5%, and 80% of these eyes with recurrent RD had type 1 hyaloid-retinal relationship (p<0.001). Nearly half the patients diagnosed as complete posterior vitreous detachment preoperatively were found to have incomplete posterior vitreous detachment intraoperatively. Majority of the cases with rhegmatogenous retinal detachment have some form of strong vitreoretinal adhesion. Hyaloid-retinal relationship varies with types of retinal breaks, retinal degeneration, and proliferative vitreoretinopathy. Intraoperative hyaloid-retinal relationship is frequently different from that assessed before surgery and the proposed classification may improve surgical decision making and prognostication.

Long-Time Plasma Membrane Imaging Based on a Two-Step Synergistic Cell Surface Modification Strategy.

PubMed

Jia, Hao-Ran; Wang, Hong-Yin; Yu, Zhi-Wu; Chen, Zhan; Wu, Fu-Gen

2016-03-16

Long-time stable plasma membrane imaging is difficult due to the fast cellular internalization of fluorescent dyes and the quick detachment of the dyes from the membrane. In this study, we developed a two-step synergistic cell surface modification and labeling strategy to realize long-time plasma membrane imaging. Initially, a multisite plasma membrane anchoring reagent, glycol chitosan-10% PEG2000 cholesterol-10% biotin (abbreviated as "GC-Chol-Biotin"), was incubated with cells to modify the plasma membranes with biotin groups with the assistance of the membrane anchoring ability of cholesterol moieties. Fluorescein isothiocyanate (FITC)-conjugated avidin was then introduced to achieve the fluorescence-labeled plasma membranes based on the supramolecular recognition between biotin and avidin. This strategy achieved stable plasma membrane imaging for up to 8 h without substantial internalization of the dyes, and avoided the quick fluorescence loss caused by the detachment of dyes from plasma membranes. We have also demonstrated that the imaging performance of our staining strategy far surpassed that of current commercial plasma membrane imaging reagents such as DiD and CellMask. Furthermore, the photodynamic damage of plasma membranes caused by a photosensitizer, Chlorin e6 (Ce6), was tracked in real time for 5 h during continuous laser irradiation. Plasma membrane behaviors including cell shrinkage, membrane blebbing, and plasma membrane vesiculation could be dynamically recorded. Therefore, the imaging strategy developed in this work may provide a novel platform to investigate plasma membrane behaviors over a relatively long time period.

Nisin and lysostaphin activity against preformed biofilm of Staphylococcus aureus involved in bovine mastitis.

PubMed

Ceotto-Vigoder, H; Marques, S L S; Santos, I N S; Alves, M D B; Barrias, E S; Potter, A; Alviano, D S; Bastos, M C F

2016-07-01

The biofilm produced by Staphylococcus aureus isolates involved in clinical or subclinical bovine mastitis and the activity of nisin and lysostaphin against the preformed biofilm produced by these strains were investigated. Eighteen strains were tested and all produced biofilm. Eight strains with distinct biofilm composition were selected for the antimicrobial activity assays. The minimal inhibitory concentration of each bacteriocin was determined against the planktonic cells and ranged from 15·6 to 500 μg ml(-1) for nisin, and from 3·9 to 50 μg ml(-1) , for lysostaphin. Lysostaphin treatment (0·4 μg ml(-1) ) for 4 h caused a strong Staph. aureus 4181 biofilm detachment and death of the majority of the sessile cells, while nisin treatment (100 μg ml(-1) ) for the same time caused only a great reduction in cell viability. Additionally, combination of both bacteriocins for 4 h resulted in significant death of the sessile cells but no biofilm detachment. The treatment with lysostaphin alone or in combination with nisin was effective in killing most biofilm sessile cells. The action of lysostaphin, either alone or in combination with nisin, against established staphylococcal biofilm may represent an alternative to bovine mastitis control. However, the duration of the treatment should be considered for its application so that the best effectiveness can be achieved. © 2016 The Society for Applied Microbiology.

Detached-Eddy Simulations of Attached and Detached Boundary Layers

NASA Astrophysics Data System (ADS)

Caruelle, B.; Ducros, F.

2003-12-01

This article presents Detached-Eddy Simulations (DESs) of attached and detached turbulent boundary layers. This hybrid Reynolds Averaged Navier-Stokes (RANS) / Large Eddy Simulation (LES) model goes continuously from RANS to LES according to the mesh definition. We propose a parametric study of the model over two "academic" configurations, in order to get information on the influence of the mesh to correctly treat complex flow with attached and detached boundary layers.

Observed use of automatic seat belts in 1987 cars.

PubMed

Williams, A F; Wells, J K; Lund, A K; Teed, N

1989-10-01

Usage of the automatic belt systems supplied by six large-volume automobile manufacturers to meet the federal requirements for automatic restraints were observed in suburban Washington, D.C., Chicago, Los Angeles, and Philadelphia. The different belt systems studied were: Ford and Toyota (motorized, nondetachable automatic shoulder belt), Nissan (motorized, detachable shoulder belt), VW and Chrysler (nonmotorized, detachable shoulder belt), and GM (nonmotorized detachable lap and shoulder belt). Use of automatic belts was significantly greater than manual belt use in otherwise comparable late-model cars for all manufacturers except Chrysler; in Chrysler cars, automatic belt use was significantly lower than manual belt use. The automatic shoulder belts provided by Ford, Nissan, Toyota, and VW increased use rates to about 90%. Because use rates were lower in Ford cars with manual belts, their increase was greater. GM cars had the smallest increase in use rates; however, lap belt use was highest in GM cars. The other manufacturers supply knee bolsters to supplement shoulder belt protection; all--except VW--also provide manual lap belts, which were used by about half of those who used the automatic shoulder belt. The results indicate that some manufacturers have been more successful than others in providing automatic belt systems that result in high use that, in turn, will mean fewer deaths and injuries in those cars.

Vitreous advanced glycation endproducts and α-dicarbonyls in retinal detachment patients with type 2 diabetes mellitus and non-diabetic controls

PubMed Central

Mulder, Douwe J.; Schalkwijk, Casper G.; Scheijen, Jean L.; Smit, Andries J.; Los, Leonoor I.

2017-01-01

Purpose Advanced glycation endproducts (AGEs) and their precursors α-dicarbonyls are implicated in the progression of diabetic retinopathy. The purpose of this study was to assess AGEs and α-dicarbonyls in the vitreous of patients with type 2 diabetes mellitus (T2DM) with early stages or absence of diabetic retinopathy. Methods We examined vitreous samples obtained during vitrectomy from 31 T2DM patients presenting themselves with rhegmatogenous retinal detachment and compared these to 62 non-diabetic rhegmatogenous retinal detachment patients, matched on age, estimated glomerular filtration rate, smoking, intra-ocular lens implantation, and proliferative vitreoretinopathy. AGEs (pentosidine, Nε-(carboxymethyl)lysine, Nε-(carboxyethyl)lysine, and 5-hydro-5-methylimidazolone) and α-dicarbonyls (3-deoxyglucosone, methylglyoxal, and glyoxal) were measured by ultra performance liquid chromatography or high performance liquid chromatography. Skin autofluorescence was measured by the AGE Reader. Results Mean age was 64 ± 7.6 years for T2DM patients and 63 ± 8.1 years for controls. For T2DM patients, median diabetes duration was 2.2 (0.3–7.4) years. Non-proliferative diabetic retinopathy was present in 1 patient and classified as absent or background retinopathy in 30 patients. Vitreous levels of pentosidine (2.20 vs. 1.59 μmol/mol lysine, p = 0.012) and 3-deoxyglucosone (809 vs. 615 nmol/L, p = 0.001) were significantly elevated in T2DM patients compared to controls. Other AGEs and α-dicarbonyls in the vitreous were not significantly different. There was a trend for increased skin autofluorescence in T2DM patients as compared to controls (p = 0.07). Conclusions Pentosidine and 3-deoxyglucosone concentrations were increased in the vitreous of rhegmatogenous retinal detachment patients with a relatively short duration of diabetes compared to non-diabetic rhegmatogenous retinal detachment patients. PMID:28264049

Investigation of plasma behavior during noble gas injection in the end-cell of GAMMA 10/PDX by using the multi-fluid code ‘LINDA’

NASA Astrophysics Data System (ADS)

Islam, M. S.; Nakashima, Y.; Hatayama, A.

2017-12-01

The linear divertor analysis with fluid model (LINDA) code has been developed in order to simulate plasma behavior in the end-cell of linear fusion device GAMMA 10/PDX. This paper presents the basic structure and simulated results of the LINDA code. The atomic processes of hydrogen and impurities have been included in the present model in order to investigate energy loss processes and mechanism of plasma detachment. A comparison among Ar, Kr and Xe shows that Xe is the most effective gas on the reduction of electron and ion temperature. Xe injection leads to strong reduction in the temperature of electron and ion. The energy loss terms for both the electron and the ion are enhanced significantly during Xe injection. It is shown that the major energy loss channels for ion and electron are charge-exchange loss and radiative power loss of the radiator gas, respectively. These outcomes indicate that Xe injection in the plasma edge region is effective for reducing plasma energy and generating detached plasma in linear device GAMMA 10/PDX.

Extending metabolome coverage for untargeted metabolite profiling of adherent cultured hepatic cells.

PubMed

García-Cañaveras, Juan Carlos; López, Silvia; Castell, José Vicente; Donato, M Teresa; Lahoz, Agustín

2016-02-01

MS-based metabolite profiling of adherent mammalian cells comprises several challenging steps such as metabolism quenching, cell detachment, cell disruption, metabolome extraction, and metabolite measurement. In LC-MS, the final metabolome coverage is strongly determined by the separation technique and the MS conditions used. Human liver-derived cell line HepG2 was chosen as adherent mammalian cell model to evaluate the performance of several commonly used procedures in both sample processing and LC-MS analysis. In a first phase, metabolite extraction and sample analysis were optimized in a combined manner. To this end, the extraction abilities of five different solvents (or combinations) were assessed by comparing the number and the levels of the metabolites comprised in each extract. Three different chromatographic methods were selected for metabolites separation. A HILIC-based method which was set to specifically separate polar metabolites and two RP-based methods focused on lipidome and wide-ranging metabolite detection, respectively. With regard to metabolite measurement, a Q-ToF instrument operating in both ESI (+) and ESI (-) was used for unbiased extract analysis. Once metabolite extraction and analysis conditions were set up, the influence of cell harvesting on metabolome coverage was also evaluated. Therefore, different protocols for cell detachment (trypsinization or scraping) and metabolism quenching were compared. This study confirmed the inconvenience of trypsinization as a harvesting technique, and the importance of using complementary extraction solvents to extend metabolome coverage, minimizing interferences and maximizing detection, thanks to the use of dedicated analytical conditions through the combination of HILIC and RP separations. The proposed workflow allowed the detection of over 300 identified metabolites from highly polar compounds to a wide range of lipids.

HPW-RX40 restores anoikis sensitivity of human breast cancer cells by inhibiting integrin/FAK signaling

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

Chen, I-Hua; Shih, Hsin-Chu; Hsieh, Pei-Wen

Anoikis is defined as apoptosis, which is induced by inappropriate cell–matrix interactions. Cancer cells with anoikis resistance tend to undergo metastasis, and this phenomenon has been reported to be associated with integrin and FAK activity. HPW-RX40 is a derivative of 3,4-methylenedioxy-β-nitrostyrene, which is known to prevent platelet aggregation by inhibition of integrin. In the present study, we investigated the effect of HPW-RX40 on an anoikis-resistant human breast cancer cell line MDA-MB-231. HPW-RX40 inhibited cell aggregation and induced cell death in suspending MDA-MB-231 cells, but had only little effect on the monolayer growth of adherent cells. Analysis of caspase activation andmore » poly (ADP-ribose) polymerase (PARP) cleavage confirmed anoikis in HPW-RX40-treated suspending cancer cells. HPW-RX40 also affected the Bcl-2 family proteins in detached cancer cells. Furthermore, HPW-RX40 inhibited detachment-induced activation of FAK and the downstream phosphorylation of Src and paxillin, but did not affect this pathway in adherent cancer cells. We also found that the expression and activation of β1 integrin in MDA-MB-231 cells were reduced by HPW-RX40. The combination of HPW-RX40 with an EGFR inhibitor led to enhanced anoikis and inhibition of the FAK pathway in breast cancer cells. Taken together, our results suggest that HPW-RX40 restores the anoikis sensitivity in the metastatic breast cancer cells by inhibiting integrin and subsequent FAK activation, and reveal a potential strategy for prevention of tumor metastasis. - Highlights: • The β-nitrostyrene derivative, HPW-RX40, induces anoikis in human breast cancer cells. • HPW-RX40 inhibits the integrin/FAK signaling pathway. • The combination of HPW-RX40 with an EGFR inhibitor leads to enhanced anoikis. • HPW-RX40 may have a potential to prevent the spread of metastatic breast cancer.« less

Cell-to-cell signaling through light: just a ghost of chance?

PubMed Central

2013-01-01

Despite the large number of reports attributing the signaling between detached cell cultures to the electromagnetic phenomena, almost no report so far included a rigorous analysis of the possibility of such signaling. In this paper, we examine the physical feasibility of the electromagnetic communication between cells, especially through light, with regard to the ambient noise illumination. We compare theoretically attainable parameters of communication with experimentally obtained data of the photon emission from cells without a specially pronounced ability of bioluminescence. We show that the weak intensity of the emission together with an unfavorable signal-to-noise ratio, which is typical for natural conditions, represent an important obstacle to the signal detection by cells. PMID:24219796

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

Covele, Brent; Kotschenreuther, M.; Mahajan, S.

The X-Divertor geometry on DIII-D has demonstrated reduced particle and heat fluxes to the target, facilitating detachment onset at ~20% lower upstream density and higher H-mode pedestal pressure than a standard divertor. SOLPS modeling suggests that this effect cannot be explained by an increase in total connection length alone, but rather by the addition of connection length specifically in the power-dissipating volume near the target, via poloidal flux expansion and flaring. But, poloidal flaring must work synergistically with divertor closure to most effectively reduce the detachment density threshold. Furthermore, the model also points to carbon radiation as the primary drivermore » of power dissipation in divertors on the DIII-D floor, which is consistent with experimental observations. Sustainable divertor detachment at lower density has beneficial consequences for energy confinement and current drive efficiency in the core for advanced tokamak (AT) operation, while simultaneously satisfying the exhaust requirements of the plasma-facing components.« less

Clinical Outcomes of Retinal Detachment Surgery following Cytomegalovirus Retinitis in Patients on Highly Active Anti-retroviral Therapy for Acquired Immune Deficiency Syndrome.

PubMed

Mathur, Gaurav; Ratra, Dhanashree; Bhuibhar, Sagar Sudhakar; Roy, Rupak

2015-01-01

The objective of this study is to describe the surgical outcomes of patients of HIV on HAART who underwent surgery for CMV retinitis related retinal detachment. A retrospective analysis of the medical records of 40 eyes of 35 consecutive HIV positive patients who underwent surgical repair for CMV retinitis associated rhegmatogenous retinal detachment between January 2000 to August 2010 was done. All patients had an adequate follow up of atleast 6 months. Favourable anatomical outcome was achieved in 78 % of eyes with the eyes having a attached retina, clear media and controlled intraocular pressure.Favourable functional outcome (vision >3/60) was achieved in 56%. Though anatomical outcomes have not changed from the pre HAART era but there has been an increase in favorable functional outcomes possibly due to effects of antiretroviral therapy.

Effects of Tacrolimus or Sirolimus on the adhesion of vascular wall cells: Controlled in-vitro comparison study.

PubMed

Krüger-Genge, A; Hiebl, B; Franke, R P; Lendlein, A; Jung, F

2017-01-01

In drug eluting stents the cytostatic drugs Sirolimus or Tacrolimus are used to inhibit blood vessel restenosis by limiting the proliferation of smooth muscle cells. However, the cytostatic activity of both drugs was shown to be not cell specific and could also affect the stent endothelialisation, respectively. Currently, only limited in vitro data are available about the impact of Sirolimus and Tacrolimus on endothelial cell proliferation over a broad concentration range. To answer this question the following study was performed.Commercially obtained HUVEC were expanded with DMEM cell culture medium (GIBCO, Germany) supplemented with 5 vol% fetal calf serum on non-coated regular polystyrene-based 24-multiwell plates. For drug testings 2×104 cells/cm2 were seeded and grown for 24 h until 30-40% of the multiwell surfaces were covered and then exposed to Sirolimus (1.0×10-11 - 1.0×10-5 mol/l) or Tacrolimus (2.0×10-8 - 6.2×10-5 mol/l), both dissolved in DMSO. 12, 24 and 48 h after adding the drugs cell numbers per area were quantified by counting the cells in six wells with four fields of view per well, representing 0.6 mm2, using a confocal laser microscope.After 48 h of cell growth in the drug-free cell culture medium, the HUVEC number increased from 2.0×104 to 3.55×104 cells/cm2 (mean cell doubling time: 53.6 h, n = 6). At lower concentrations (≤2.0×10-6 mol/l) Tacrolimus reduced the number of adherent HUVEC significantly less than Sirolimus (p < 0.05). However, at higher concentrations (≥2.07×10-5 mol/l) the effect of Tacrolimus on the number of adherent endothelial cells was significantly greater than that of Sirolimus (p < 0.05). At the highest concentration applied (6.22×10-5 mol/l), Tacrolimus induced detachment of all HUVECs within 12 h after drug application. The number of adherent HUVEC decreased only slightly (about 9%) after Sirolimus application at the highest concentration (1.09×10-5 mol/l).These data show that in a non-flow model the cytostatic drug Tacrolimus reduced the number of adherent endothelial cells less than Sirolimus, as long as the drug concentration did not surpass 10-6 mol/l. At the limits of solubility, Sirolimus (1×10-5 mol/l) reduced the number of adherent endothelial cells less than Tacrolimus (6×10-5 mol/l), which induced detachment of endothelial cells.

Ozone injury increases infection of geranium leaves by Botrytis cinerea

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

Manning, W.J.; Feder, W.A.; Perkins, I.

1970-04-01

Detached and attached, inoculated and noninoculated, ozone-injured and noninjured leaves from the lower, middle, and terminal regions of plants of geranium cultivars Enchantress and White Mountain were observed for infection by Botrytis cinerea. Previous exposure to ozone did not appreciably influence the susceptibility of leaves of either geranium cultivar to infection by B. cinerea, unless there was visible ozone injury. Ozone-injured, necrotic tissues on older attached and detached geranium leaves of both cultivars served as infection courts for B. cinerea. 14 references, 1 table.

Modeling of Detached Solidification

NASA Technical Reports Server (NTRS)

Regel, Liya L.; Wilcox, William R.; Popov, Dmitri

1997-01-01

Our long term goal is to develop techniques to achieve detached solidification reliably and reproducibly, in order to produce crystals with fewer defects. To achieve this goal it is necessary to understand thoroughly the physics of detached solidification. It was the primary objective of the current project to make progress toward this complete understanding. 'Me products of this grant are attached. These include 4 papers and a preliminary survey of the observations of detached solidification in space. We have successfully modeled steady state detached solidification, examined the stability of detachment, and determined the influence of buoyancy-driven convection under different conditions. Directional solidification in microgravity has often led to ingots that grew with little or no contact with the ampoule wall. When this occurred, crystallographic perfection was usually greatly improved -- often by several orders of magnitude. Indeed, under the Soviet microgravity program the major objective was to achieve detached solidification with its resulting improvement in perfection and properties. Unfortunately, until recently the true mechanisms underlying detached solidification were unknown. As a consequence, flight experiments yielded erratic results. Within the past three years, we have developed a new theoretical model that explains many of the flight results. This model gives rise to predictions of the conditions required to yield detached solidification.

Chitosan as an adjuvant-like substrate for dendritic cell culture to enhance antitumor effects.

PubMed

Lin, Yong-Chong; Lou, Pei-Jen; Young, Tai-Horng

2014-10-01

To induce monocyte differentiation into dendritic cells (DCs) is the essential protocol for the DC-mediated cancer immunotherapy. In this study, monocytes isolated from mouse bone marrow were cultured on chitosan substrate to evaluate the effect of the chitosan culture system on the induction and tumor protection of DCs. Compared to tissue culture polystyrene (TCPS), the chitosan culture system could enhance monocyte aggregation and detachment with increased MTT reduction activity and expression of DC marker CD11c and LPS co-receptor CD14. Moreover, compared to TCPS, chitosan could enhance lipopolysaccharides (LPS)-stimulated DCs to secrete higher amount of IL-12. More importantly, vaccination of tumor lysate-pulsed DCs harvested from chitosan could increase cytotoxic T-lymphocyte (CTL) activity and showed significantly enhanced anti-tumor effect than those from TCPS. Therefore, the current study demonstrated that a protocol to culture DCs on a less-adherent chitosan substrate followed by treatment with tumor lysate has the potential in future DC-based vaccine application. Copyright © 2014 Elsevier Ltd. All rights reserved.

RSV-encoded NS2 promotes epithelial cell shedding and distal airway obstruction

PubMed Central

Liesman, Rachael M.; Buchholz, Ursula J.; Luongo, Cindy L.; Yang, Lijuan; Proia, Alan D.; DeVincenzo, John P.; Collins, Peter L.; Pickles, Raymond J.

2014-01-01

Respiratory syncytial virus (RSV) infection is the major cause of bronchiolitis in young children. The factors that contribute to the increased propensity of RSV-induced distal airway disease compared with other commonly encountered respiratory viruses remain unclear. Here, we identified the RSV-encoded nonstructural 2 (NS2) protein as a viral genetic determinant for initiating RSV-induced distal airway obstruction. Infection of human cartilaginous airway epithelium (HAE) and a hamster model of disease with recombinant respiratory viruses revealed that NS2 promotes shedding of infected epithelial cells, resulting in two consequences of virus infection. First, epithelial cell shedding accelerated the reduction of virus titers, presumably by clearing virus-infected cells from airway mucosa. Second, epithelial cells shedding into the narrow-diameter bronchiolar airway lumens resulted in rapid accumulation of detached, pleomorphic epithelial cells, leading to acute distal airway obstruction. Together, these data indicate that RSV infection of the airway epithelium, via the action of NS2, promotes epithelial cell shedding, which not only accelerates viral clearance but also contributes to acute obstruction of the distal airways. Our results identify RSV NS2 as a contributing factor for the enhanced propensity of RSV to cause severe airway disease in young children and suggest NS2 as a potential therapeutic target for reducing the severity of distal airway disease. PMID:24713657

HEMOXCell, a New Oxygen Carrier Usable as an Additive for Mesenchymal Stem Cell Culture in Platelet Lysate-Supplemented Media.

PubMed

Le Pape, Fiona; Cosnuau-Kemmat, Lucie; Richard, Gaëlle; Dubrana, Frédéric; Férec, Claude; Zal, Franck; Leize, Elisabeth; Delépine, Pascal

2017-04-01

Human mesenchymal stem cells (MSCs) are promising candidates for therapeutic applications such as tissue engineering. However, one of the main challenges is to improve oxygen supply to hypoxic areas to reduce oxygen gradient formation while preserving MSC differentiation potential and viability. For this purpose, a marine hemoglobin, HEMOXCell, was evaluated as an oxygen carrier for culturing human bone marrow MSCs in vitro for future three-dimensional culture applications. Impact of HEMOXCell on cell growth and viability was assessed in human platelet lysate (hPL)-supplemented media. Maintenance of MSC features, such as multipotency and expression of MSC specific markers, was further investigated by biochemical assays and flow cytometry analysis. Our experimental results highlight its oxygenator potential and indicate that an optimal concentration of 0.025 g/L HEMOXCell induces a 25%-increase of the cell growth rate, preserves MSC phenotype, and maintains MSC differentiation properties; a two-fold higher concentration induces cell detachment without altering cell viability. Our data suggest the potential interest of HEMOXCell as a natural oxygen carrier for tissue engineering applications to oxygenate hypoxic areas and to maintain cell viability, functions and "stemness." These features will be further tested within three-dimensional scaffolds. © 2017 International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

Efficient adhesion-based plasma membrane isolation for cell surface N-glycan analysis.

PubMed

Mun, Ji-Young; Lee, Kyung Jin; Seo, Hoon; Sung, Min-Sun; Cho, Yee Sook; Lee, Seung-Goo; Kwon, Ohsuk; Oh, Doo-Byoung

2013-08-06

Glycans, which decorate cell surfaces, play crucial roles in various physiological events involving cell surface recognition. Despite the importance of surface glycans, most analyses have been performed using total cells or whole membranes rather than plasma membranes due to difficulties related to isolation. In the present study, we employed an adhesion-based method for plasma membrane isolation to analyze N-glycans on cell surfaces. Cells were attached to polylysine-coated glass plates and then ruptured by hypotonic pressure. After washing to remove intracellular organelles, only a plasma membrane fraction remained attached to the plates, as confirmed by fluorescence imaging using organelle-specific probes. The plate was directly treated with trypsin to digest and detach the glycoproteins from the plasma membrane. From the resulting glycopeptides, N-glycans were released and analyzed using MALDI-TOF mass spectrometry and HPLC. When N-glycan profiles obtained by this method were compared to those by other methods, the amount of high-mannose type glycans mainly contaminated from the endoplasmic reticulum was dramatically reduced, which enabled the efficient detection of complex type glycans present on the cell surface. Moreover, this method was successfully used to analyze the increase of high-mannose glycans on the surface as induced by a mannosidase inhibitor treatment.

Detached Bridgman Growth of Germanium and Germanium-Silicon Alloy Crystals

NASA Technical Reports Server (NTRS)

Szofran, F. R.; Volz, M. P.; Schweizer, M.; Cobb, S. D.; Motakef, S.; Croell, A.; Dold, P.; Curreri, Peter A. (Technical Monitor)

2002-01-01

Earth based experiments on the science of detached crystal growth are being conducted on germanium and germanium-silicon alloys (2 at% Si average composition) in preparation for a series of experiments aboard the International Space Station (ISS). The purpose of the microgravity experiments includes differentiating among proposed mechanisms contributing to detachment, and confirming or refining our understanding of the detachment mechanism. Because large contact angle are critical to detachment, sessile drop measurements were used to determine the contact angles as a function of temperature and composition for a large number of substrates made of potential ampoule materials. Growth experiments have used pyrolytic boron nitride (pBN) and fused silica ampoules with the majority of the detached results occurring predictably in the pBN. The contact angles were 173 deg (Ge) and 165 deg (GeSi) for pBN. For fused silica, the contact angle decreases from 150 deg to an equilibrium value of 117 deg (Ge) or from 129 deg to an equilibrium value of 100 deg (GeSi) over the duration of the experiment. The nature and extent of detachment is determined by using profilometry in conjunction with optical and electron microscopy. The stability of detachment has been analyzed, and an empirical model for the conditions necessary to achieve sufficient stability to maintain detached growth for extended periods has been developed. Results in this presentation will show that we have established the effects on detachment of ampoule material, pressure difference above and below the melt, and silicon concentration; samples that are nearly completely detached can be grown repeatedly in pBN.

A cell impedance measurement device for the cytotoxicity assay dependent on the velocity of supplied toxic fluid

NASA Astrophysics Data System (ADS)

Kang, Yoon-Tae; Kim, Min-Ji; Cho, Young-Ho

2018-04-01

We present a cell impedance measurement chip capable of characterizing the toxic response of cells depending on the velocity of the supplied toxic fluid. Previous impedance-based devices using a single open-top chamber have been limited to maintaining a constant supply velocity, and devices with a single closed-top chamber present difficulties in simultaneous cytotoxicity assay for varying levels of supply velocities. The present device, capable of generating constant and multiple levels of toxic fluid velocity simultaneously within a single stepwise microchannel, performs a cytotoxicity assay dependent on toxic fluid velocity, in order to find the effective velocity of toxic fluid to cells for maximizing the cytotoxic effect. We analyze the cellular toxic response of 5% ethanol media supplied to cancer cells within a toxic fluid velocity range of 0-8.3 mm s-1. We observe the velocity-dependent cell detachment rate, impedance, and death rate. We find that the cell detachment rate decreased suddenly to 2.4% at a velocity of 4.4 mm s-1, and that the change rates of cell resistance and cell capacitance showed steep decreases to 8% and 41%, respectively, at a velocity of 5.7 mm s-1. The cell death rate and impedance fell steeply to 32% at a velocity of 5.7 mm s-1. We conclude that: (1) the present device is useful in deciding on the toxic fluid velocity effective to cytotoxicity assay, since the cellular toxic response is dependent on the velocity of toxic fluid, and; (2) the cell impedance analysis facilitates a finer cellular response analysis, showing better correlation with the cell death rate, compared to conventional visual observation. The present device, capable of performing the combinational analysis of toxic fluid velocity and cell impedance, has potential for application to the fine cellular toxicity assay of drugs with proper toxic fluid velocity.

32 CFR 700.723 - Administration and discipline: Separate and detached command.

Code of Federal Regulations, 2010 CFR

2010-07-01

... detached command. 700.723 Section 700.723 National Defense Department of Defense (Continued) DEPARTMENT OF... Administration and discipline: Separate and detached command. Any flag or general officer in command, any officer... are separate or detached commands. Such officer shall state in writing that it is a separate or...

32 CFR 700.723 - Administration and discipline: Separate and detached command.

Code of Federal Regulations, 2014 CFR

2014-07-01

... detached command. 700.723 Section 700.723 National Defense Department of Defense (Continued) DEPARTMENT OF... Administration and discipline: Separate and detached command. Any flag or general officer in command, any officer... are separate or detached commands. Such officer shall state in writing that it is a separate or...

32 CFR 700.723 - Administration and discipline: Separate and detached command.

Code of Federal Regulations, 2011 CFR

2011-07-01

... detached command. 700.723 Section 700.723 National Defense Department of Defense (Continued) DEPARTMENT OF... Administration and discipline: Separate and detached command. Any flag or general officer in command, any officer... are separate or detached commands. Such officer shall state in writing that it is a separate or...

32 CFR 700.723 - Administration and discipline: Separate and detached command.

Code of Federal Regulations, 2013 CFR

2013-07-01

... detached command. 700.723 Section 700.723 National Defense Department of Defense (Continued) DEPARTMENT OF... Administration and discipline: Separate and detached command. Any flag or general officer in command, any officer... are separate or detached commands. Such officer shall state in writing that it is a separate or...

32 CFR 700.723 - Administration and discipline: Separate and detached command.

Code of Federal Regulations, 2012 CFR

2012-07-01

... detached command. 700.723 Section 700.723 National Defense Department of Defense (Continued) DEPARTMENT OF... Administration and discipline: Separate and detached command. Any flag or general officer in command, any officer... are separate or detached commands. Such officer shall state in writing that it is a separate or...

Erosion: Irrigation-induced

USDA-ARS?s Scientific Manuscript database

Soil can be eroded by sprinkler or surface irrigation. Once sprinkler droplet kinetic energy detaches soil, overland flow transports the sediment downslope and off-site. Protecting the soil surface, increasing sprinkler wetted diameters, and tilling to increase infiltration and thereby lessen overla...

Exposure of Bacterial Biofilms to Electrical Current Leads to Cell Death Mediated in Part by Reactive Oxygen Species.

PubMed

Brinkman, Cassandra L; Schmidt-Malan, Suzannah M; Karau, Melissa J; Greenwood-Quaintance, Kerryl; Hassett, Daniel J; Mandrekar, Jayawant N; Patel, Robin

2016-01-01

Bacterial biofilms may form on indwelling medical devices such as prosthetic joints, heart valves and catheters, causing challenging-to-treat infections. We have previously described the 'electricidal effect', in which bacterial biofilms are decreased following exposure to direct electrical current. Herein, we sought to determine if the decreased bacterial quantities are due to detachment of biofilms or cell death and to investigate the role that reactive oxygen species (ROS) play in the observed effect. Using confocal and electron microscopy and flow cytometry, we found that direct current (DC) leads to cell death and changes in the architecture of biofilms formed by Gram-positive and Gram-negative bacteria. Reactive oxygen species (ROS) appear to play a role in DC-associated cell death, as there was an increase in ROS-production by Staphylococcus aureus and Staphylococcus epidermidis biofilms following exposure to DC. An increase in the production of ROS response enzymes catalase and superoxide dismutase (SOD) was observed for S. aureus, S. epidermidis and Pseudomonas aeruginosa biofilms following exposure to DC. Additionally, biofilms were protected from cell death when supplemented with antioxidants and oxidant scavengers, including catalase, mannitol and Tempol. Knocking out SOD (sodAB) in P. aeruginosa led to an enhanced DC effect. Microarray analysis of P. aeruginosa PAO1 showed transcriptional changes in genes related to the stress response and cell death. In conclusion, the electricidal effect results in death of bacteria in biofilms, mediated, at least in part, by production of ROS.

Localization and Quantification of Callose in the Streptophyte Green Algae Zygnema and Klebsormidium: Correlation with Desiccation Tolerance

PubMed Central

Herburger, Klaus; Holzinger, Andreas

2015-01-01

Freshwater green algae started to colonize terrestrial habitats about 460 million years ago, giving rise to the evolution of land plants. Today, several streptophyte green algae occur in aero-terrestrial habitats with unpredictable fluctuations in water availability, serving as ideal models for investigating desiccation tolerance. We tested the hypothesis that callose, a β-d-1,3-glucan, is incorporated specifically in strained areas of the cell wall due to cellular water loss, implicating a contribution to desiccation tolerance. In the early diverging genus Klebsormidium, callose was drastically increased already after 30 min of desiccation stress. Localization studies demonstrated an increase in callose in the undulating cross cell walls during cellular water loss, allowing a regulated shrinkage and expansion after rehydration. This correlates with a high desiccation tolerance demonstrated by a full recovery of the photosynthetic yield visualized at the subcellular level by Imaging-PAM. Furthermore, abundant callose in terminal cell walls might facilitate cell detachment to release dispersal units. In contrast, in the late diverging Zygnema, the callose content did not change upon desiccation for up to 3.5 h and was primarily localized in the corners between individual cells and at terminal cells. While these callose deposits still imply reduction of mechanical damage, the photosynthetic yield did not recover fully in the investigated young cultures of Zygnema upon rehydration. The abundance and specific localization of callose correlates with the higher desiccation tolerance in Klebsormidium when compared with Zygnema. PMID:26412780

Bacteriophage PRD1 batch experiments to study attachment, detachment and inactivation processes

NASA Astrophysics Data System (ADS)

Sadeghi, Gholamreza; Schijven, Jack F.; Behrends, Thilo; Hassanizadeh, S. Majid; van Genuchten, Martinus Th.

2013-09-01

Knowledge of virus removal in subsurface environments is pivotal for assessing the risk of viral contamination of water resources and developing appropriate protection measures. Columns packed with sand are frequently used to quantify attachment, detachment and inactivation rates of viruses. Since column transport experiments are very laborious, a common alternative is to perform batch experiments where usually one or two measurements are done assuming equilibrium is reached. It is also possible to perform kinetic batch experiments. In that case, however, it is necessary to monitor changes in the concentration with time. This means that kinetic batch experiments will be almost as laborious as column experiments. Moreover, attachment and detachment rate coefficients derived from batch experiments may differ from those determined using column experiments. The aim of this study was to determine the utility of kinetic batch experiments and investigate the effects of different designs of the batch experiments on estimated attachment, detachment and inactivation rate coefficients. The experiments involved various combinations of container size, sand-water ratio, and mixing method (i.e., rolling or tumbling by pivoting the tubes around their horizontal or vertical axes, respectively). Batch experiments were conducted with clean quartz sand, water at pH 7 and ionic strength of 20 mM, and using the bacteriophage PRD1 as a model virus. Values of attachment, detachment and inactivation rate coefficients were found by fitting an analytical solution of the kinetic model equations to the data. Attachment rate coefficients were found to be systematically higher under tumbling than under rolling conditions because of better mixing and more efficient contact of phages with the surfaces of the sand grains. In both mixing methods, more sand in the container yielded higher attachment rate coefficients. A linear increase in the detachment rate coefficient was observed with increased solid-water ratio using tumbling method. Given the differences in the attachment rate coefficients, and assuming the same sticking efficiencies since chemical conditions of the batch and column experiments were the same, our results show that collision efficiencies of batch experiments are not the same as those of column experiments. Upscaling of the attachment rate from batch to column experiments hence requires proper understanding of the mixing conditions. Because batch experiments, in which the kinetics are monitored, are as laborious as column experiments, there seems to be no major advantage in performing batch instead of column experiments.

Management of inadvertent needle penetration resulting in subretinal triamcinolone acetonide and retinal detachment.

PubMed

Tran, Kimberly D; Crane, Ashley M; Flynn, Harry W

2018-06-01

To report management of inadvertent needle penetration during subtenons triamcinolone acetonide administration resulting in retinal detachment. A 71-year-old female with history of diabetes, hypothyroidism, and mild myopia underwent subtenons triamcinolone acetonide (TA) injection in the right eye for nodular scleritis. There was unexpected patient movement concurrent with the injection resulting in needle penetration, subretinal and intravitreal injection of TA, superotemporal retinal break, and macula-involving retinal detachment. The patient underwent partial subretinal TA removal, successful retinal detachment repair, and recovered 20/25 visual acuity. In spite of prominent subretinal TA and retinal detachment, successful repair of retinal detachment and recovery of good visual acuity is possible.

A history of detachable coils: 1987-2012.

PubMed

Hui, Ferdinand K; Fiorella, David; Masaryk, Thomas J; Rasmussen, Peter A; Dion, Jacques E

2014-03-01

The development of detachable coils is one of the most pivotal developments in neurointervention, providing a tool that could be used to treat a wide variety of hemorrhagic stroke. From the original Guglielmi detachable coil, a number of different coil designs and delivery designs have evolved. This article reviews the history of commercially available detachable coils. A timeline of detachable coils was constructed and coil design philosophies were reviewed. A complete list of commercially available coils is presented in a timeline format. Detachable coil technology continues to evolve. Advances in construction and design have yielded products which may benefit patients in terms of safety, radiation dose reduction and cost of treatment. Continued evolution is expected, irrespective of competing disruptive technologies.

The Relationship between Job Demands and Employees' Counterproductive Work Behaviors: The Mediating Effect of Psychological Detachment and Job Anxiety.

PubMed

Chen, Yang; Li, Shuang; Xia, Qing; He, Chao

2017-01-01

This study aims to explore the relation between job demands and counterproductive work behaviors (CWBs). A cross-sectional sample of 439 coal miners completed a self-report questionnaire that assessed their job demands, psychological detachment, job anxiety, and CWBs in a Chinese context. The conceptual model, based on the stressor-detachment model, was examined using structural equation modeling. The results suggest that psychological detachment mediates not only the relation between job demands and job anxiety but also that between job demands and CWBs. Furthermore, the relation between job demands and CWBs is sequentially mediated by psychological detachment and job anxiety. Our findings validate the effectiveness of the stressor-detachment model. Moreover, we demonstrate that the underlying mechanism of the relation between job demands and CWBs can be explained by psychological detachment and job anxiety.

The Relationship between Job Demands and Employees’ Counterproductive Work Behaviors: The Mediating Effect of Psychological Detachment and Job Anxiety

PubMed Central

Chen, Yang; Li, Shuang; Xia, Qing; He, Chao

2017-01-01

This study aims to explore the relation between job demands and counterproductive work behaviors (CWBs). A cross-sectional sample of 439 coal miners completed a self-report questionnaire that assessed their job demands, psychological detachment, job anxiety, and CWBs in a Chinese context. The conceptual model, based on the stressor-detachment model, was examined using structural equation modeling. The results suggest that psychological detachment mediates not only the relation between job demands and job anxiety but also that between job demands and CWBs. Furthermore, the relation between job demands and CWBs is sequentially mediated by psychological detachment and job anxiety. Our findings validate the effectiveness of the stressor-detachment model. Moreover, we demonstrate that the underlying mechanism of the relation between job demands and CWBs can be explained by psychological detachment and job anxiety. PMID:29163274

Otoancorin Knockout Mice Reveal Inertia is the Force for Hearing

NASA Astrophysics Data System (ADS)

Weddell, Thomas; Legan, P. Kevin; Lukashkina, Victoria A.; Goodyear, Richard J.; Welstead, Lindsy; Petit, Chistine; Russell, Ian J.; Lukashkin, Andrei N.; Richardson, Guy P.

2011-11-01

We demonstrate that in Otoa-/- mice, in which the inner-ear-specific protein otoancorin is absent, excitation of the outer hair cells and cochlear amplification is normal. This finding is remarkable because the tectorial membrane (TM), although remaining functionally attached to the outer hair cell bundles, is completely detached from the spiral limbus. Therefore, as in ancestral vertebrate auditory organs, where inertia provides the excitatory force to the hair cells, it is the inertia of the TM that must be important for exciting the outer hair cells, setting the sensitivity of their transducer conductance, and determining the precise timing of cochlear amplification.

Ultrasonographic study on changes in axial eye dimensions after encircling procedure in retinal detachment surgery.

PubMed

Larsen, J S; Syrdalen, P

1979-06-01

The pre- and postoperative results of ultrasonographic measurements on the axial ocular components in 10 phakic eyes with retinal detachment treated with encircling silicone rubber band are presented. A significant increase (P less than 0.001) in axial eye length from 0.62 to 1.24 mm (average: 0.98 mm) was found. The elongation of the eye was caused by a corresponding increase in the length of the vitreous cavity. No significant changes were found in the anterior segment of the eyes. These data demonstrate that the postoperative refractive change in a myopic direction, which an encircling procedure with moderate indentation often produces, is caused by an axial elongation of the eye.

Key factors of eddy current separation for recovering aluminum from crushed e-waste.

PubMed

Ruan, Jujun; Dong, Lipeng; Zheng, Jie; Zhang, Tao; Huang, Mingzhi; Xu, Zhenming

2017-02-01

Recovery of e-waste in China had caused serious pollutions. Eddy current separation is an environment-friendly technology of separating nonferrous metallic particles from crushed e-waste. However, due to complex particle characters, separation efficiency of traditional eddy current separator was low. In production, controllable operation factors of eddy current separation are feeding speed, (ωR-v), and S p . There is little special information about influencing mechanism and critical parameters of these factors in eddy current separation. This paper provided the special information of these key factors in eddy current separation of recovering aluminum particles from crushed waste refrigerator cabinets. Detachment angles increased as the increase of (ωR-v). Separation efficiency increased with the growing of detachment angles. Aluminum particles were completely separated from plastic particles in critical parameters of feeding speed 0.5m/s and detachment angles greater than 6.61deg. S p /S m of aluminum particles in crushed waste refrigerators ranged from 0.08 to 0.51. Separation efficiency increased as the increase of S p /S m . This enlightened us to develop new separator to separate smaller nonferrous metallic particles in e-waste recovery. High feeding speed destroyed separation efficiency. However, greater S p of aluminum particles brought positive impact on separation efficiency. Greater S p could increase critical feeding speed to offer greater throughput of eddy current separation. This paper will guide eddy current separation in production of recovering nonferrous metals from crushed e-waste. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effect of pH on bacteriophage transport through sandy soils

USGS Publications Warehouse

Kinoshita, Takashi; Bales, Roger C.; Maguire, Kimberley M.; Gerba, Charles P.

1993-01-01

Effects of pH and hydrophobicity on attachment and detachment of PRD-1 and MS-2 in three different sandy soils were investigated in a series of laboratory-column experiments. Concentrations of the lipid-containing phage PRD-1 decreased 3–4 orders of magnitude during passage through the 10–15-cm-long columns. Attachment of the lipid-containing phage PRD-1 was insensitive to pH and was apparently controlled by hydrophobic interactions in soil media. The less-hydrophobic phage MS-2 acted conservatively; it was not removed in the columns at pH's 5.7–8.0. The sticking efficiency (α) in a colloid-filtration model was between 0.1 and 1 for PRD-1, indicating a relatively high removal efficiency. Phage attachment was reversible, but detachment under steady-state conditions was slow. An increase in pH had a moderate effect on enhancing detachment. Still, these soils should continue to release phage to virus-free water for days to weeks following exposure to virus-containing water. In sandy soils with a mass-fraction organic carbon as low as a few hundredths of a percent, pH changes in the range 5.7–8.0 should have little effect on retention of more-hydrophobic virus (e.g., PRD-1), in that retardation will be dominated by hydrophobic effects. Sharp increases in pH should enhance detachment and transport of virus previously deposited on soil grains. A more hydrophilic virus (e.g., MS-2) will transport as a conservative tracer in low-carbon sandy soil.

Atomistic investigation on the detachment of oil molecules from defective alumina surface

NASA Astrophysics Data System (ADS)

Xie, W. K.; Sun, Y. Z.; Liu, H. T.

2017-12-01

The mechanism of oil detachment from defective alumina surface in aqueous solution was investigated via atomistic molecular dynamics (MD) simulations. Special attention was focused on the effect of surface defect on the oil detachment. Our simulation results suggest that compared with perfect Al2O3 surface, defective substrate surface provides much more sites for the adsorption of oil molecules, thus it has higher oil adsorption energy. However, higher oil-solid adsorption energy does not mean that oil contaminants are much more difficult to be detached. It is found that surface defect could induce the spontaneous imbibition of water molecules, effectively promoting the detachment of oil molecules. Thus, compared with perfect alumina surface, the detachment of oil molecules from defective alumina surface tends to be much easier. Moreover, surface defect could lead to the oil residues inside surface defect. In water solution, the entire detachment process of oil molecules on defective surface consists of following stages, including the early detachment of oil molecules inside surface defect induced by capillary-driven spontaneous imbibition of water molecules, the following conformational change of oil molecules on topmost surface and the final migration of detached oil molecules from solid surface. These findings may help to sufficiently enrich the removal mechanism of oil molecules adhered onto defective solid surface.

The study of adhesive forces between the type-3 fimbriae of Klebsiella pneumoniae and collagen-coated surfaces by using optical tweezers

NASA Astrophysics Data System (ADS)

Chan, Chiahan; Fan, Chia-chieh; Huang, Ying-Jung; Peng, Hwei-Ling; Long, Hsu

2004-10-01

Adherence to host cells by a bacterial pathogen is a critical step for establishment of infection. It will contribute greatly to the understanding of bacterial pathogenesis by studying the biological force between a single pair of pathogen and host cell. In our experiment, we use a calibrated optical tweezers system to detach a single Klebsiella pneumoniae, the pathogen, from collagen, the host. By gradually increasing the laser power of the optical tweezers until the Klebsiella pneumoniae is detached from the collagen, we obtain the magnitude of the adhesive force between them. This happens when the adhesive force is barely equal to the trapping force provided by the optical tweezers at that specific laser power. This study is important because Klebsiella pneumoniae is an opportunistic pathogen which causes suppurative lesions, urinary and respiratory tract infections. It has been proved that type 3 fimbrial adhesin (mrkD) is strongly associated with the adherence of Klebsiella pneumoniae. Besides, four polymorphic mrkD alleles: namely, mrkDv1, v2, v3, and v4, are typed by using RFLP. In order to investigate the relationship between the structure and the function for each of these variants, DNA fragments encoding the major fimbrial proteins mrkA, mrkB, mrkC are expressed together with any of the four mrkD adhesins in E. coli JM109. Our study shows that the E. coli strain carrying the mrkDv3 fimbriae has the strongest binding activity. This suggests that mrkDv3 is a key factor that enhances the adherence of Klebsiella Pneumoniae to human body.

Alterations in leucocyte subsets and histomorphology in normal-appearing perilesional skin and early and chronic hidradenitis suppurativa lesions.

PubMed

van der Zee, H H; de Ruiter, L; Boer, J; van den Broecke, D G; den Hollander, J C; Laman, J D; Prens, E P

2012-01-01

Current insight into the histopathological course of events during disease progression in hidradenitis suppurativa (HS) is fragmentary. To identify histological alterations and leucocyte subsets in normal-appearing perilesional skin, and early and chronic HS lesions. In this observational study we examined eight perilesional skin samples, and six early and 10 chronic prototypic HS lesions, as well as skin samples from four healthy donors using in situ immunostaining. Perilesional skin showed mild psoriasiform hyperplasia and follicular plugging as well as a low-grade influx of tryptase-positive mast cells, CD3+ T cells, CD138+ plasma cells and factor XIIIa+ dendritic cells. In early HS lesions, neutrophilic abscess formation and influx of mainly macrophages, monocytes and dendritic cells predominated. In chronic disease, the infiltrate expanded with markedly increased frequencies of CD20+ and CD79a+ B cells and CD138+ plasma cells. As in early lesions, free keratin fibres were detected in the dermis and within giant cells. Single detached keratinocytes and strands of follicular epithelium were observed in the dermis, the latter frequently expressing Ki67, indicative of active proliferation. Psoriasiform hyperplasia, follicular plugging and low-grade leucocytic infiltration are already present in normal-appearing perilesional skin. Keratin fibres in the dermis are associated with clinical disease. Early lesions are characterized by neutrophilic abscess formation and influx of mainly histiocytes, and chronic lesions mainly by expansion of B cells and plasma cells in 'pseudo' follicles. Proliferating strands of follicular epithelium may initiate fistula formation. Mast cells are increased in all stages of HS including perilesional skin. © 2011 The Authors. BJD © 2011 British Association of Dermatologists.

Final Act of an Oceanic Detachment Fault Revealed by Submersible Dives at 13°48'N on the Mid-Atlantic Ridge

NASA Astrophysics Data System (ADS)

Parnell-Turner, R. E.; Mittelstaedt, E. L.; Kurz, M. D.; Klein, F.

2017-12-01

A large proportion of crustal accretion on the slow-spreading Mid-Atlantic Ridge occurs under the influence of slip on low-angle detachment faults. The final stages of activity on an individual detachment system remain poorly understood, since it is difficult to place age constraints on exposed fault surfaces or lava flows. We use data from a combination of manned (Alvin) and autonomous (Sentry) submersible dives on a detachment near 13°48'N, to infer the history of slip and volcanism on a detachment fault which has recently become extinct. The corrugated surface, near the toe of the detachment, is cross-cut by a volcanic ridge, where pillow lavas have been photographed and sampled. Sub-bottom (CHIRP) profiles acquired by Sentry provide estimates of sediment thickness, which we use as a proxy for seafloor age, thus providing a relative dating tool for the exposed detachment footwall and erupted lavas. Sediments covering the footwall are 2 m thinner than those on lavas which cut across the detachment, implying that slip continued for 150 ka after eruption (assuming a constant sedimentation rate of 7 ± 2 mm/yr). Alternatively, sediment on the footwall may have been mass-wasted, and volcanism could have been contemporaneous with detachment inactivity. These results demonstrate that detachment faults may be highly sensitive to local changes in magma supply, and that direct seafloor observations are crucial to understanding slow-spreading ridge mechanics.

The e[sup [minus

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

Holroyd, R.A.; Schwarz, H.A.; Stradowska, E.

The rate constants for attachment of excess electrons to 1,3-butadiene (k[sub a]) and detachment from the butadiene anion (k[sub d]) in n-hexane are reported. The equilibrium constant, K[sub eq] = k[sub a]/k[sub d], increases rapidly with pressure and decreases as the temperature increases. At -7[degree]C attachment is observed at 1 bar. At high pressures the attachment rate is diffusion controlled. The activation energy for detachment is about 21 kcal/mol; detachment is facilitated by the large entropy of activation. The reaction volumes for attachment range from -181 cm[sup 3]/mol at 400 bar to-122 cm[sup 3]/mol at 1500 bar and are largelymore » attributed to the electrostriction volume of the butadiene anion ([Delta][bar V][sub el]). Values of [Delta][bar V][sub el] calculated by a model, which includes a glassy shell of solvent molecules around the ion, are in agreement with experimental reaction volumes. The analysis indicates the partial molar volume of the electron in hexane is small and probably negative. It is shown that the entropies of reaction are closely related to the partial molar volumes of reaction. 22 refs., 5 figs., 5 tabs.« less

Regulation of Cell Cytoskeleton and Membrane Mechanics by Electric Field: Role of Linker Proteins

PubMed Central

Titushkin, Igor; Cho, Michael

2009-01-01

Abstract Cellular mechanics is known to play an important role in the cell homeostasis including proliferation, motility, and differentiation. Significant variation in the mechanical properties between different cell types suggests that control of the cell metabolism is feasible through manipulation of the cell mechanical parameters using external physical stimuli. We investigated the electrocoupling mechanisms of cellular biomechanics modulation by an electrical stimulation in two mechanically distinct cell types—human mesenchymal stem cells and osteoblasts. Application of a 2 V/cm direct current electric field resulted in approximately a twofold decrease in the cell elasticity and depleted intracellular ATP. Reduction in the ATP level led to inhibition of the linker proteins that are known to physically couple the cell membrane and cytoskeleton. The membrane separation from the cytoskeleton was confirmed by up to a twofold increase in the membrane tether length that was extracted from the cell membrane after an electrical stimulation. In comparison to human mesenchymal stem cells, the membrane-cytoskeleton attachment in osteoblasts was much stronger but, in response to the same electrical stimulation, the membrane detachment from the cytoskeleton was found to be more pronounced. The observed effects mediated by an electric field are cell type- and serum-dependent and can potentially be used for electrically assisted cell manipulation. An in-depth understanding and control of the mechanisms to regulate cell mechanics by external physical stimulus (e.g., electric field) may have great implications for stem cell-based tissue engineering and regenerative medicine. PMID:19167316

Hydrogen sulfide-powered solid oxide fuel cells

NASA Astrophysics Data System (ADS)

Liu, Man

2004-12-01

The potential utilization of hydrogen sulfide as fuel in solid oxide fuel cells has been investigated using an oxide-ion conducting YSZ electrolyte and different kinds of anode catalysts at operating temperatures in the range of 700--900°C and at atmospheric pressure. This technology offers an economically attractive alternative to present methods for removing toxic and corrosive H2S gas from sour gas streams and a promising approach for cogenerating electrical energy and useful chemicals. The primary objective of the present research was to find active and stable anode materials. Fuel cell experimental results showed that platinum was a good electrocatalyst for the conversion of H2S, but the Pt/YSZ interface was physically unstable due to the reversible formation and decomposition of PtS in H 2S streams at elevated temperatures. Moreover, instability of the Pt/YSZ interface was accelerated significantly by electrochemical reactions, and ultimately led to the detachment of the Pt anode from the electrolyte. It has been shown that an interlayer of TiO2 stabilized the Pt anode on YSZ electrolyte, thereby prolonging cell lifetime. However, the current output for a fuel cell using Pt/TiO2 as anode was not improved compared to using Pt alone. It was therefore necessary to investigate novel anode systems for H 2S-air SOFCs. New anode catalysts comprising composite metal sulfides were developed. These catalysts exhibited good electrical conductivity and better catalytic activity than Pt. In contrast to MoS2 alone, composite catalysts (M-Mo-S, M = Fe, Co, Ni) were not volatile and had superior stability. However, when used for extended periods of time, detachment of Pt current collecting film from anodes comprising metal sulfides alone resulted in a large increase in contact resistance and reduction in cell performance. Consequently, a systematic investigation was conducted to identify alternative electronic conductors for use with M-Mo-S catalysts. Anode catalysts comprising Co-Mo-S admixed with up to 10% Ag powder were found to have excellent performance and longevity, as well as improved electrical contact when compared with Pt/M-Mo-S anode systems. The highest current density of 450 mA/cm2 and power density of 115 mW/cm2 were achieved with an anode that consisted of 95% (Co-Mo-S) and 5% Ag.

[Screening and identification of anoikis-resistant gene UBCH7 in esophageal cancer cells].

PubMed

Yang, Yang; Wang, Bo-Shi; Wang, Xiao-Min; Zhang, Yu; Wang, Ming-Rong; Jia, Xue-Mei

2012-02-01

Anoikis is a kind of programmed cell death induced by loss of extracellular matrix (ECM) adhesion, which is one of key factors for homestasis. Resistance to anoikis is required for tumor cell metastasis. We have previously shown several anoikis-resistance genes in esophageal squamous cell carcinoma (ESCC). In order to find novel anoikis-resistant genes in ESCC, we constructed retroviral cDNA library using total RNA from ESCC cell lines. NIH 3T3 cells, which are sensitive to anoikis, were infected with the library constructed. The cells were cultured in soft agar, and the clones which can survive in detached states were selected. The cDNAs inserted into the anoikis-resistant NIH3T3 clones were amplified using retroviral specific primers. Sequencing analysis showed that a cDNA fragment inserted into the anoikis-resistant clone contains full coding sequence (ORF) of human UBCH7/UBE2L3 gene. By infection with retrovirus encoding UBCH7 ORF (pMSCV-UBCH7), forced expression of UBCH7 increased the anoikis-resistance of NIH3T3 cells. More importantly, knockdown of UBCH7 expression by siRNA transfection reduced the anoikis-resistant ability of esophageal cancer MLuC1 cells. The data suggest that UBCH7/UBE2L3 gene would be involved in anoikis-resistance in ESCC.

Smart polymeric nanoparticles with pH-responsive and PEG-detachable properties for co-delivering paclitaxel and survivin siRNA to enhance antitumor outcomes.

PubMed

Jin, Mingji; Jin, Guangming; Kang, Lin; Chen, Liqing; Gao, Zhonggao; Huang, Wei

2018-01-01

The co-delivery of chemotherapeutic agents and small interfering RNA (siRNA) within one cargo can enhance the anticancer outcomes through its synergistic therapeutic effects. We prepared smart polymeric nanoparticles (NPs) with pH-responsive and poly(ethylene glycol) (PEG)-detachable properties to systemically co-deliver paclitaxel (PTX) and siRNA against survivin gene for lung cancer therapy. The cationic polyethyleneimine-block-polylactic acid (PEI-PLA) was first synthesized and characterized, with good biocompatibility. PTX was encapsulated into the hydrophobic core of the PEI-PLA polymers by dialysis, and then the survivin siRNA was loaded onto the PTX-loaded NPs (PEI-PLA/PTX) through electrostatic interaction between siRNA and PEI block. Finally, the negatively charged poly(ethylene glycol)-block-poly(L-aspartic acid sodium salt) (PEG-PAsp) was coated onto the surface of NPs by electrostatic interaction to form final smart polymeric NPs with mean particle size of 82.4 nm and zeta potential of 4.1 mV. After uptake of NPs by tumor cells, the PEG-PAsp segments became electrically neutral owing to the lower endosome pH and consequently detached from the smart NPs. This process allowed endosomal escape of the NPs through the proton-sponge effect of the exposed PEI moiety. The resulting NPs achieved drug loading of 6.04 wt% and exhibited good dispersibility within 24 h in 10% fetal bovine serum (FBS). At pH 5.5, the NPs presented better drug release and cellular uptake than at pH 7.4. The NPs with survivin siRNA effectively knocked down the expression of survivin mRNA and protein owing to enhanced cell uptake of NPs. Cell counting kit-8 (CCK-8) assay showed that the NPs presented low systemic toxicity and improved antiproliferation effect of PTX on A549 cells. Moreover, in vivo studies demonstrated that accumulated NPs in the tumor site were capable of inhibiting the tumor growth and extending the survival rate of the mice by silencing the survivin gene and delivering PTX into tumor cells simultaneously. These results indicate that the prepared nano-vectors could be a promising co-delivery system for novel chemo/gene combination therapy.

New Insights into PhaM-PhaC-Mediated Localization of Polyhydroxybutyrate Granules in Ralstonia eutropha H16

PubMed Central

Bresan, Stephanie

2017-01-01

ABSTRACT The formation and localization of polyhydroxybutyrate (PHB) granules in Ralstonia eutropha are controlled by PhaM, which interacts both with the PHB synthase (PhaC) and with the bacterial nucleoid. Here, we studied the importance of proline and lysine residues of two C-terminal PAKKA motifs in PhaM for their importance in attaching PHB granules to DNA by in vitro and in vivo methods. Substitution of the lysine residues but not of the proline residues resulted in detachment of formed PHB granules from the nucleoid. Instead, formation of PHB granule clusters at polar regions of the rod-shaped cells and an unequal distribution of PHB granules to daughter cells were observed. The formation of PHB granules was studied by the expression of chromosomally anchored gene fusions of fluorescent proteins with PhaM and PhaC in different backgrounds. PhaM and PhaC fusions showed a distinct colocalization at formed PHB granules in the nucleoid region of the wild type. In a ΔphaC background, PhaM and the catalytically inactive PhaCC319A protein were not able to form fluorescent foci, indicating that correct positioning requires the formation of PHB. Furthermore, time-lapse experiments revealed that PhaC and PhaM proteins detach from formed PHB granules at later stages, resulting in a nonhomogeneous population of PHB granules. This could explain why growth of individual PHB granules stops under PHB-permissive conditions at a certain size. IMPORTANCE PHB granules are storage compounds for carbon and energy in many prokaryotes. Equal distribution of accumulated PHB granules during cell division is therefore important for optimal fitness of the daughter cells. In R. eutropha, PhaM is responsible for maximal activity of PHB synthase, for initiation of PHB granule formation at discrete regions in the cells, and for association of formed PHB granules with the nucleoid. Here we found that four lysine residues of C-terminal PhaM sequence motifs are essential for association of PHB granules with the nucleoid. Furthermore, we followed PHB granule formation by time-lapse microscopy and provide evidence for aging of PHB granules that is manifested by detachment of previously PHB granule-associated PhaM and PHB synthase. PMID:28389545

Smart polymeric nanoparticles with pH-responsive and PEG-detachable properties for co-delivering paclitaxel and survivin siRNA to enhance antitumor outcomes

PubMed Central

Jin, Mingji; Jin, Guangming; Kang, Lin; Chen, Liqing; Gao, Zhonggao; Huang, Wei

2018-01-01

Background The co-delivery of chemotherapeutic agents and small interfering RNA (siRNA) within one cargo can enhance the anticancer outcomes through its synergistic therapeutic effects. Materials and methods We prepared smart polymeric nanoparticles (NPs) with pH-responsive and poly(ethylene glycol) (PEG)-detachable properties to systemically co-deliver paclitaxel (PTX) and siRNA against survivin gene for lung cancer therapy. The cationic polyethyleneimine-block-polylactic acid (PEI-PLA) was first synthesized and characterized, with good biocompatibility. PTX was encapsulated into the hydrophobic core of the PEI-PLA polymers by dialysis, and then the survivin siRNA was loaded onto the PTX-loaded NPs (PEI-PLA/PTX) through electrostatic interaction between siRNA and PEI block. Finally, the negatively charged poly(ethylene glycol)-block-poly(L-aspartic acid sodium salt) (PEG-PAsp) was coated onto the surface of NPs by electrostatic interaction to form final smart polymeric NPs with mean particle size of 82.4 nm and zeta potential of 4.1 mV. After uptake of NPs by tumor cells, the PEG-PAsp segments became electrically neutral owing to the lower endosome pH and consequently detached from the smart NPs. This process allowed endosomal escape of the NPs through the proton-sponge effect of the exposed PEI moiety. Results The resulting NPs achieved drug loading of 6.04 wt% and exhibited good dispersibility within 24 h in 10% fetal bovine serum (FBS). At pH 5.5, the NPs presented better drug release and cellular uptake than at pH 7.4. The NPs with survivin siRNA effectively knocked down the expression of survivin mRNA and protein owing to enhanced cell uptake of NPs. Cell counting kit-8 (CCK-8) assay showed that the NPs presented low systemic toxicity and improved antiproliferation effect of PTX on A549 cells. Moreover, in vivo studies demonstrated that accumulated NPs in the tumor site were capable of inhibiting the tumor growth and extending the survival rate of the mice by silencing the survivin gene and delivering PTX into tumor cells simultaneously. Conclusion These results indicate that the prepared nano-vectors could be a promising co-delivery system for novel chemo/gene combination therapy. PMID:29719390

Promotion of malignant phenotype after disruption of the three-dimensional structure of cultured spheroids from colorectal cancer

PubMed Central

Endo, Hiroko; Ono, Hiromasa; Hagihara, Takeshi; Okuyama, Hiroaki; Nishizawa, Yasuko; Tomita, Yasuhiko; Ohue, Masayuki; Okita, Kouki; Oyama, Hidejiro; Bono, Hidemasa; Masuko, Takashi; Inoue, Masahiro

2018-01-01

Individual and small clusters of cancer cells may detach from the edges of a main tumor and invade vessels, which can act as the origin of metastasis; however, the mechanism for this phenomenon is not well understood. Using cancer tissue-originated spheroids, we studied whether disturbing the 3D architecture of cancer spheroids can provoke the reformation process and progression of malignancy. We developed a mechanical disruption method to achieve homogenous disruption of the spheroids while maintaining cell–cell contact. After the disruption, 9 spheroid lines from 9 patient samples reformed within a few hours, and 3 of the 9 lines exhibited accelerated spheroid growth. Marker expression, spheroid forming capacity, and tumorigenesis indicated that stemness increased after spheroid disruption. In addition, the spheroid forming capacity increased in 6 of 11 spheroid lines. The disruption signature determined by gene expression profiling supported the incidence of remodeling and predicted the prognosis of patients with colorectal cancer. Furthermore, WNT and HER3 signaling were increased in the reformed spheroids, and suppression of these signaling pathways attenuated the increased proliferation and stemness after the disruption. Overall, the disruption and subsequent reformation of cancer spheroids promoted malignancy-related phenotypes through the activation of the WNT and ERBB pathways. PMID:29662620

Analytical method for determining rill detachment rate of purple soil as compared with that of loess soil

USDA-ARS?s Scientific Manuscript database

Rill detachment is an important process in rill erosion. The rill detachment rate is the fundamental basis for determination of the parameters of a rill erosion model. In this paper, an analytical method was proposed to estimate the rill detachment rate. The method is based on the exact analytical s...

Vitreous vascular endothelial growth factor concentrations in proliferative diabetic retinopathy versus proliferative vitreoretinopathy.

PubMed

Citirik, Mehmet; Kabatas, Emrah Utku; Batman, Cosar; Akin, Kadir Okhan; Kabatas, Naciye

2012-01-01

To assess vitreous vascular endothelial growth factor (VEGF) concentrations in proliferative diabetic retinopathy (PDR) in comparison to proliferative vitreoretinopathy (PVR). Vitreous samples were collected from 69 eyes of 69 patients with traumatic lens dislocation (n = 10), grade B PVR with rhegmatogenous retinal detachment (n = 13), grade C PVR with rhegmatogenous retinal detachment (n = 14), PDR with vitreous hemorrhage (n = 18), and PDR with vitreous hemorrhage and tractional retinal detachment (n = 14). Vitreous fluid samples were obtained at vitrectomy, and the levels of VEGF were measured by enzyme-linked immunosorbent assay. The mean vitreous level of VEGF was 15.14 ± 5.22 pg/ml in eyes with grade B PVR, 99.15 ± 38.58 pg/ml in eyes with grade C PVR, 4,534.01 ± 1,193.28 pg/ml in eyes with vitreous hemorrhage secondary to PDR, 5,157.29 ± 969.44 pg/ml in eyes with vitreous hemorrhage and tractional retinal detachment secondary to PDR, and 16.19 ± 5.76 pg/ml in eyes of the control group with traumatic lens dislocation. Vitreous VEGF concentrations were significantly higher in the patients with grade C PVR, PDR with vitreous hemorrhage and PDR with vitreous hemorrhage and tractional retinal detachment in comparison to the control patients (p < 0.05). A significant alteration was not observed in patients with grade B PVR (p = 0.55). Vitreous VEGF concentrations are increased in PDR and grade C PVR. The high VEGF concentrations could suggest a possible effect of VEGF on advanced PVR. Copyright © 2011 S. Karger AG, Basel.

Efficacy of two different thiol-modified crosslinked hyaluronate formulations as vitreous replacement compared to silicone oil in a model of retinal detachment

PubMed Central

Schnichels, Sven; Schneider, Nele; Hohenadl, Christine; Hurst, José; Schatz, Andreas; Januschowski, Kai; Spitzer, Martin S.

2017-01-01

The efficacy of two novel artificial vitreous body substitutes (VBS) consisting of highly biocompatible thiolated cross-linked hyaluronic acid (HA)-based hydrogels in comparison to silicone oil in a model of retinal detachment was investigated. Pars plana vitrectomy (23G) was performed in the right eye of 24 pigmented rabbits. Retinal detachment of two quadrants was induced by creating a small retinotomy near the vascular arcade and injecting balanced salt solution (BSS) subretinally. The retina was reattached by injecting air, which was followed by increasing the infusion pressure, and the retinal tear was treated by endolaser photocoagulation. At the end of the procedure, the eye was filled either with 5000-cs silicone oil (after fluid air exchange) or the respective hydrogel (with two different viscosities). Follow-up examination included slit lamp examination, funduscopy, intraocular pressure measurements (IOP), optical coherence tomography (OCT) and electroretinogram (ERG) measurements. After a maximum follow-up of four weeks both eyes were removed, examined macroscopically, photographed, and prepared for histology. Of the eight rabbits that received silicone oil, seven (87.5%) developed a recurrent retinal detachment with pronounced proliferative vitreoretinopathy within the first two weeks after surgery. In contrast, in the hydrogel treated eyes, the retina stayed attached in the majority of the cases (73.3%). IOP and retinal morphology were normal as long as the retina remained re-attached. In conclusions, this model of retinal detachment, both thiolated crosslinked hyaluronate hydrogels showed superior efficacy when compared to silicone oil. These hydrogels have a promising potential as novel vitreous body substitutes. PMID:28248989

Repair of Traumatic Rhegmatogenous Retinal Detachment Combined with Congenital Falciform Retinal Detachment.

PubMed

Mano, Fukutaro; Chang, Kuo-Chung; Mano, Tomiya

2018-01-01

To report a case of surgical repair of traumatic rhegmatogenous retinal detachment combined with congenital falciform retinal detachment (FRD). A retrospective case report. A 36-year-old man with traumatic rhegmatogenous retinal detachment complicating a previously known FRD was successfully treated despite residual FRD following pars plana lensectomy, vitrectomy, and encircling scleral buckling. His best corrected visual acuity improved from hand motion at 50 cm to 20/1,000. We concluded that the root of the FRD is susceptible to trauma because of the contraction of fibrovascular tissue. The early intervention of modern vitrectomy to traumatic rhegmatogenous retinal detachment complicating a previously known FRD is an important consideration for enhanced quality of care and optimal patient outcomes.

Repair of Traumatic Rhegmatogenous Retinal Detachment Combined with Congenital Falciform Retinal Detachment

PubMed Central

Mano, Fukutaro; Chang, Kuo-Chung; Mano, Tomiya

2018-01-01

Purpose To report a case of surgical repair of traumatic rhegmatogenous retinal detachment combined with congenital falciform retinal detachment (FRD). Methods A retrospective case report. Results A 36-year-old man with traumatic rhegmatogenous retinal detachment complicating a previously known FRD was successfully treated despite residual FRD following pars plana lensectomy, vitrectomy, and encircling scleral buckling. His best corrected visual acuity improved from hand motion at 50 cm to 20/1,000. Conclusion We concluded that the root of the FRD is susceptible to trauma because of the contraction of fibrovascular tissue. The early intervention of modern vitrectomy to traumatic rhegmatogenous retinal detachment complicating a previously known FRD is an important consideration for enhanced quality of care and optimal patient outcomes. PMID:29643782

Ocular toxoplasmosis and retinal detachment: five case reports.

PubMed

Kianersi, F; Naderi Beni, A; Ghanbari, H; Fazel, F

2012-10-01

Ocular toxoplasmosis is a potentially blinding cause of posterior uveitis. Retinal detachment is rare complication of ocular toxoplasmosis. To report the clinical course and prognosis of retinal breaks and detachments occurring in patients with ocular toxoplasmosis. This study was a retrospective, non-comparative case series of five patients with ocular toxoplasmosis who had consulted us with retinal detachment. All of the participants had retinal detachment after severe and treatment resistant toxoplasmic retinochoroiditis, leaving one of them with decreased visual acuity to light perception in spite of treatment and final visual acuity was 20/100 or better in four patients. The functional prognosis for the patients with retinal detachment was poor. Careful retinal examination in ocular toxoplasmosis is warranted, especially in patients with severe intraocular inflammation.

The role of detachment faulting in slow seafloor spreading: First results from cruise JC132 to the MAR at 13N

NASA Astrophysics Data System (ADS)

Reston, T. J.; Falder, M.; Peirce, C.; Simão, N.; Searle, R. C.; MacLeod, C. J.

2016-12-01

Our understanding of the processes of seafloor spreading at slow-spreading mid-ocean ridges is undergoing a paradigm shift as it has become increasingly clear that much of the slowly spread seafloor has not been built solely by the symmetric accretion of the products of partial melting to the trailing edges of the separating plates, but that tectonic stretching on large-offset normal (`detachment') faults, weakened by the penetration of water and production of weak phyllosilicates (e.g. talc), also plays a fundamental role, unroofing plutonic and partially serpentinized mantle footwalls to form `oceanic core complexes' (OCCs). However, fundamental aspects of OCC structure and evolution, and the detachment process itself, are still poorly understood: it is not clear, and consequently hotly debated, whether the controlling faults are seismically active, what their 3D geometry is, how they are linked with the supply and emplacement of magma and, crucially, how far detachments continue laterally in the sub-surface and/or if they link at depth. Our understanding of the 3D geometry and mechanics of detachment faults is limited by a paucity of observations from actively forming OCCs, and in particular the lack of sub-surface images from which the geometry and extent of surface features can be traced to depth, the lack of detailed P- and S-wave 3D-volume velocity models to reveal how detachments relate to magmatic accretion in time and space, and the lack of coincident observations of local seismicity to determine the focus of active deformation. To investigate these key questions, in Jan-Feb 2016 we collected a combined MCS - wide-angle seismic dataset, and high resolution near surface and near seafloor bathymetry and magnetics, to complement passive recordings of microseismicity made in 2014. We discuss the aims of the cruise, and present first results.

Structural evolution and mechanical behaviour of Pt nanoparticle superlattices at high pressure

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

Zhu, Jinlong; Quan, Zewei; Wang, Chenyu

High pressure is an effective means for tuning the interparticle distances of nanoparticle (NP) superlattice and thus for modifying their physical properties and functionalities. In this work, we determined the evolutio of inter-NP distances of a Pt NP superlattice with increasing pressure using an in situ synchrotro small-angle X-ray scattering (SAXS) technique in a diamond-anvil cell (DAC). Transmission electro microscopy (TEM) was used to characterize the microstructures of pre- and post-compression samples Our results demonstrate that the evolution of Pt NP assemblies with increasing pressure consists of fou stages: (1) ligand elastic response, (2) uniform compression, (3) ligand detachment frommore » NP surfaces, an (4) deviatoric compression of ligands between neighboring NPs. Lastly, by controlling the magnitudes of applie pressure and deviatoric stress, one can sinter NPs into novel architectures such as nanowires an nanoceramics.« less

Structural evolution and mechanical behaviour of Pt nanoparticle superlattices at high pressure

DOE PAGES

Zhu, Jinlong; Quan, Zewei; Wang, Chenyu; ...

2016-02-05

High pressure is an effective means for tuning the interparticle distances of nanoparticle (NP) superlattice and thus for modifying their physical properties and functionalities. In this work, we determined the evolutio of inter-NP distances of a Pt NP superlattice with increasing pressure using an in situ synchrotro small-angle X-ray scattering (SAXS) technique in a diamond-anvil cell (DAC). Transmission electro microscopy (TEM) was used to characterize the microstructures of pre- and post-compression samples Our results demonstrate that the evolution of Pt NP assemblies with increasing pressure consists of fou stages: (1) ligand elastic response, (2) uniform compression, (3) ligand detachment frommore » NP surfaces, an (4) deviatoric compression of ligands between neighboring NPs. Lastly, by controlling the magnitudes of applie pressure and deviatoric stress, one can sinter NPs into novel architectures such as nanowires an nanoceramics.« less

Effects of silk fibroin in murine dry eye

NASA Astrophysics Data System (ADS)

Kim, Chae Eun; Lee, Ji Hyun; Yeon, Yeung Kyu; Park, Chan Hum; Yang, Jaewook

2017-03-01

The study aimed to investigate the effects of silk fibroin in a mouse model of dry eye. The experimental dry eye mouse model was developed using more than twelve-weeks-old NOD.B10.H2b mice exposing them to 30-40% ambient humidity and injecting them with scopolamine hydrobromide for 10 days. Tear production and corneal irregularity score were measured by the instillation of phosphate buffered saline or silk fibroin. Corneal detachment and conjunctival goblet cell density were observed by hematoxylin and eosin or periodic acid Schiff staining in the cornea or conjunctiva. The expression of inflammatory markers was detected by immunohistochemistry in the lacrimal gland. The silk group tear production was increased, and corneal smoothness was improved. The corneal epithelial cells and conjunctival goblet cells were recovered in the silk groups. The expression of inflammatory factors was inhibited in the lacrimal gland of the silk group. These results show that silk fibroin improved the cornea, conjunctiva, and lacrimal gland in the mouse model of dry eye. These findings suggest that silk fibroin has anti-inflammatory effects in the experimental models of dry eye.

Divertor heat flux mitigation in the National Spherical Torus Experimenta)

NASA Astrophysics Data System (ADS)

Soukhanovskii, V. A.; Maingi, R.; Gates, D. A.; Menard, J. E.; Paul, S. F.; Raman, R.; Roquemore, A. L.; Bell, M. G.; Bell, R. E.; Boedo, J. A.; Bush, C. E.; Kaita, R.; Kugel, H. W.; Leblanc, B. P.; Mueller, D.; NSTX Team

2009-02-01

Steady-state handling of divertor heat flux is a critical issue for both ITER and spherical torus-based devices with compact high power density divertors. Significant reduction of heat flux to the divertor plate has been achieved simultaneously with favorable core and pedestal confinement and stability properties in a highly shaped lower single null configuration in the National Spherical Torus Experiment (NSTX) [M. Ono et al., Nucl. Fusion 40, 557 2000] using high magnetic flux expansion at the divertor strike point and the radiative divertor technique. A partial detachment of the outer strike point was achieved with divertor deuterium injection leading to peak flux reduction from 4-6MWm-2to0.5-2MWm-2 in small-ELM 0.8-1.0MA, 4-6MW neutral beam injection-heated H-mode discharges. A self-consistent picture of the outer strike point partial detachment was evident from divertor heat flux profiles and recombination, particle flux and neutral pressure measurements. Analytic scrape-off layer parallel transport models were used for interpretation of NSTX detachment experiments. The modeling showed that the observed peak heat flux reduction and detachment are possible with high radiated power and momentum loss fractions, achievable with divertor gas injection, and nearly impossible to achieve with main electron density, divertor neutral density or recombination increases alone.

The interplay between cytokinins and light during senescence in detached Arabidopsis leaves.

PubMed

Janečková, Helena; Husičková, Alexandra; Ferretti, Ursula; Prčina, Maroš; Pilařová, Eva; Plačková, Lenka; Pospíšil, Pavel; Doležal, Karel; Špundová, Martina

2018-05-09

Light and cytokinins are known to be the key players in the regulation of plant senescence. In detached leaves, the retarding effect of light on senescence is well described; however, it is not clear to what extent is this effect connected with changes in endogenous cytokinin levels. We have performed a detailed analysis of changes in endogenous content of 29 cytokinin forms in detached leaves of Arabidopsis thaliana (wild-type and three cytokinin receptor double-mutants). Leaves were kept under different light conditions and changes in cytokinin content were correlated with changes in chlorophyll content, efficiency of photosystem II photochemistry and lipid peroxidation. In leaves kept in darkness we have observed decreased content of the most abundant cytokinin free bases and ribosides, but the content of cis-zeatin increased, which indicates the role of this cytokinin in the maintenance of basal leaf viability. Our findings underscore the importance of light conditions on the content of specific cytokinins, especially N 6 -(Δ 2 -isopentenyl)adenine. Based on our results, we present a scheme summarizing the contribution of the main active forms of cytokinins, cytokinin receptors and light to senescence regulation. We conclude that light can compensate the disrupted cytokinin signaling in detached leaves. This article is protected by copyright. All rights reserved.

Simultaneous requirement of carbon dioxide and abscisic acid for stomatal closing in Xanthium strumarium L.

PubMed

Raschke, K

1975-01-01

Open stomata of detached leaves of Xanthium strumarium L. closed only when carbon dioxide and abscisic acid (ABA) were presented simultaneously. Three parameters of stomatal closing were determined after additions of ABA to the irrigation water of detached leaves, while the leaves were exposed to various CO2 concentrations ([CO2]s) in the air; a) the delay between addition of ABA and a reduction of stomatal conductance by 5%, b) the velocity of stomatal closing, and c) the new conductance. Changes in all three parameters showed that stomatal responses to ABA were enhanced by CO2; this effect followed saturation kinetics. Half saturation occurred at an estimated [CO2] in the stomatal pore of 200 μl l(-1). With respect to ABA, stomata responded in normal air with half their maximal amplitude at [ABA]s between 10(-6) and 10(-5) M(+-)-ABA. The amounts of ABA taken up by the leaves during the delay increased with a power <1 (on the average, 0.67) of the [ABA] in the transpiration stream. The minimal amount of ABA found to produce a stomatal response was about 1 pmol of (+-)-ABA per cm(2) leaf area, almost two orders of magnitude smaller than the original content of the leaves in ABA indicating that most of the endogenous ABA was in a compartment isolated from the guard cells.An interaction between stomatal responses to CO2 and ABA was also found in Gossypium hirsutum L. and Commelina communis L.; it was however much weaker than in X. strumarium.Based on earlier findings and on the results of this investigation it is suggested that stomata close if the cytoplasm of the guard cells contains much malate and H(+). The acid content in turn is determined by the relative rates of production of malic acid (from endogenous as well as exogenous CO2) and its removal (by transport of the anion into the vacuole and exchange of the H(+) for K(+) with the environment of the guard cells). The simultaneous requirement of CO2 and ABA for stomatal closure leads to the inference that ABA inhibits the expulsion of H(+) from guard cells.

Comparison of Sulfur Hexafluoride (SF6) and Air Tamponade in Noniridectomized Descemet Membrane Endothelial Keratoplasty.

PubMed

Einan-Lifshitz, Adi; Sorkin, Nir; Boutin, Tanguy; Showail, Mahmood; Borovik, Armand; Jamshidi, Farzad; Chan, Clara C; Rootman, David S

2018-03-01

To compare the efficacy and safety of 20% sulfur hexafluoride gas (SF6) and air tamponade in patients who underwent noniridectomized Descemet membrane endothelial keratoplasty (DMEK). A retrospective chart review of patients who underwent DMEK with either air or SF6 tamponade: 41 eyes received air tamponade (group 1) and 41 received SF6 tamponade (group 2). Best spectacle-corrected visual acuity, endothelial cell density, and complications including graft detachment and elevated intraocular pressure were compared. The mean follow-up time was 8 ± 4 months in group 1 and 3 ± 2 months in group 2. Mean best spectacle-corrected visual acuity improved from 1.12 ± 0.88 to 0.64 ± 0.78 logarithm of the minimum angle of resolution (logMAR) in group 1 (P = 0.009) and from 1.00 ± 0.78 to 0.62 ± 0.53 logMAR in group 2 (P = 0.006). The graft detachment rate was 39% (16 eyes) in group 1 and 42% (17 eyes) in group 2 (P = 0.822). The rate of graft detachment larger than one third of the graft area was 17% in group 1 and 20% in group 2 (P = 0.775). Rebubbling was performed in 26.8% and 20% of eyes in group 1 and 2, respectively (P = 0.43). Average endothelial cell loss was 32% in group 1 and 33% in group 2 (P = 0.83). In the immediate postoperative period, elevated intraocular pressure was observed in 2 eyes (5%) in group 1 and in 4 eyes (10%) in group 2 (P = 0.4). There was 1 primary graft failure in each group. Use of air with it being readily available and short acting is a good method of Descemet membrane tamponade in noniridectomized DMEK.

Fibronectin matrix assembly suppresses dispersal of glioblastoma cells.

PubMed

Sabari, Joshua; Lax, Daniel; Connors, Daniel; Brotman, Ian; Mindrebo, Eric; Butler, Christine; Entersz, Ildiko; Jia, Dongxuan; Foty, Ramsey A

2011-01-01

Glioblastoma (GBM), the most aggressive and most common form of primary brain tumor, has a median survival of 12-15 months. Surgical excision, radiation and chemotherapy are rarely curative since tumor cells broadly disperse within the brain. Preventing dispersal could be of therapeutic benefit. Previous studies have reported that increased cell-cell cohesion can markedly reduce invasion by discouraging cell detachment from the tumor mass. We have previously reported that α5β1 integrin-fibronectin interaction is a powerful mediator of indirect cell-cell cohesion and that the process of fibronectin matrix assembly (FNMA) is crucial to establishing strong bonds between cells in 3D tumor-like spheroids. Here, we explore a potential role for FNMA in preventing dispersal of GBM cells from a tumor-like mass. Using a series of GBM-derived cell lines we developed an in vitro assay to measure the dispersal velocity of aggregates on a solid substrate. Despite their similar pathologic grade, aggregates from these lines spread at markedly different rates. Spreading velocity is inversely proportional to capacity for FNMA and restoring FNMA in GBM cells markedly reduces spreading velocity by keeping cells more connected. Blocking FNMA using the 70 KDa fibronectin fragment in FNMA-restored cells rescues spreading velocity, establishing a functional role for FNMA in mediating dispersal. Collectively, the data support a functional causation between restoration of FNMA and decreased dispersal velocity. This is a first demonstration that FNMA can play a suppressive role in GBM dispersal.

Border cell release: Cell separation without cell wall degradation?

PubMed

Mravec, Jozef

2017-07-03

Plant border cells are specialized cells derived from the root cap with roles in the biomechanics of root growth and in forming a barrier against pathogens. The mechanism of highly localized cell separation which is essential for their release to the environment is little understood. Here I present in situ analysis of Brachypodium distachyon, a model organism for grasses which possess type II primary cell walls poor in pectin content. Results suggest similarity in spatial dynamics of pectic homogalacturonan during dicot and monocot border cell release. Integration of observations from different species leads to the hypothesis that this process most likely does not involve degradation of cell wall material but rather uses unique cell wall structural and compositional means enabling both the rigidity of the root cap as well as detachability of given cells on its surface.

Risk of progression in macula-on rhegmatogenous retinal detachment.

PubMed

Callizo, Josep; Pfeiffer, Sebastian; Lahme, Eva; van Oterendorp, Christian; Khattab, Mohammed; Bemme, Sebastian; Kulanga, Miroslav; Hoerauf, Hans; Feltgen, Nicolas

2017-08-01

To identify factors that may lead to a rapid progression in macula-on rhegmatogenous retinal detachment (RRD), in particular, those that may lead to macular involvement. Observational, prospective, single-center study. Patients referred for surgery due to primary rhegmatogenous retinal detachment with the macula on between 2009 and 2013 were included. Relevant factors analyzed included age, time delay until surgery, lens status, myopia, the detachment's location and configuration as well as number, size and type of retinal breaks. Eyes underwent optical coherence tomography to detect macular detachment. A multivariate analysis was performed to investigate the effect of several factors in the progression of retinal detachment. A total of 116 eyes of 116 patients were included. Mean time delay between admission and surgery was 1.8 ± 1.4 days. Progression was observed in 19.8% of the eyes. Of those, 47.8% presented macular detachment. Ten of the 11 (90.9%) eyes presenting progression involving the macula also exhibited a bullous configuration, which was the only parameter that correlated significantly with detachment progression in patients with (p = 0.0036) and without (p = 0.0014) macular involvement. For the first time in a prospective trial, a bullous configuration was found to be a highly significant predictor for progression in macula-on detachments. Our data support prompt surgery in patients diagnosed with bullous macula-on RRD.

The influence of blade profile and slots on the performance of a centrifugal impeller

NASA Astrophysics Data System (ADS)

Fowler, H. S.

1980-01-01

As part of the program of studies on centrifugal impellers, the problem of instability at low flows was investigated. The major cause was found to be flow detachment from the impeller vanes. Slotted blades were found to be the most effective means of delaying this detachment, and extending the working range of the blower. Low speed studies were confirmed by a test program on a high speed machine, where it was demonstrated that the improved flow range was accompanied by a general increase of efficiency. The design and placement of the slots is discussed.

Microbial Attachment Inhibition through Low-Voltage Electrochemical Reactions on Electrically Conducting Membranes.

PubMed

Ronen, Avner; Duan, Wenyan; Wheeldon, Ian; Walker, Sharon; Jassby, David

2015-11-03

Bacterial biofilm formation on membrane surfaces remains a serious challenge in water treatment systems. The impact of low voltages on microbial attachment to electrically conducting ultrafiltration membranes was investigated using a direct observation cross-flow membrane system mounted on a fluorescence microscope. Escherichia coli and microparticle deposition and detachment rates were measured as a function of the applied electrical potential to the membrane surface. Selecting bacteria and particles with low surface charge minimized electrostatic interactions between the bacteria and charged membrane surface. Application of an electrical potential had a significant impact on the detachment of live bacteria in comparison to dead bacteria and particles. Image analysis indicated that when a potential of 1.5 V was applied to the membrane/counter electrode pair, the percent of dead bacteria was 32±2.1 and 67±3.6% when the membrane was used as a cathode or anode, respectively, while at a potential of 1 V, 92±2.4% were alive. The application of low electrical potentials resulted in the production of low (μM) concentrations of hydrogen peroxide (HP) through the electroreduction of oxygen. The electrochemically produced HP reduced microbial cell viability and increased cellular permeability. Exposure to low concentrations of electrochemically produced HP on the membrane surface prevents bacterial attachment, thus ensuring biofilm-free conditions during membrane filtration operations.

31 CFR 358.6 - What is the procedure for converting bearer corpora and detached bearer coupons to book-entry?

Code of Federal Regulations, 2011 CFR

2011-07-01

... bearer corpora and detached bearer coupons to book-entry? 358.6 Section 358.6 Money and Finance: Treasury... PUBLIC DEBT REGULATIONS GOVERNING BOOK-ENTRY CONVERSION OF BEARER CORPORA AND DETACHED BEARER COUPONS § 358.6 What is the procedure for converting bearer corpora and detached bearer coupons to book-entry...

The Generation of Oceanic Lithosphere in an Embryonic Oceanic Crust : the Example of the Chenaillet Ophiolite in the Western Alps

NASA Astrophysics Data System (ADS)

Masini, E.; Manatschal, G.; Muntener, O.

2007-12-01

The Chenaillet Ophiolite exposed in the Franco-Italian Alps represents a well-preserved ocean-floor sequence that was only weakly affected by later Alpine convergence. Based on the similarity between rock types and structures reported from ultraslow spreading ridges and those observed in the Chenaillet Ophiolite, it may represent a field analogue for slow to ultraslow spreading ridges such as the Gakkel Ridge or the Southwest Indian Ridge. Mapping of the Chenaillet Ophiolite enabled to identify an oceanic detachment fault that extends over a surface of about 16 km2 capping exhumed mantle and gabbros onto which clastic sediments have been deposited. The footwall of the detachment is formed by mafic and ultramafic rocks. The mantle rocks are strongly serpentinized lherzolites and subordinate harzburgites and dunites. Microstructures reminiscent of impregnation, and cpx major and trace element chemistry indicate that spinel peridotite is (locally) replaced by plagioclase-bearing assemblages. Pyroxene thermometry on primary minerals indicates high temperatures of equilibration ( max 1200°C) for the mantle rocks. Gabbros range from troctolite and olivine-gabbros to Fe-Ti gabbros and show clear evidence of syn-magmatic deformation, partially obliterated by retrograde amphibolite and low-grade metamorphic conditions. In sections perpendicular to the detachment within the footwall, syn-tectonic gabbros and serpentinized peridotites grade over some tens of meters into cataclasites that are capped by fault gouges. Petro-structural investigations of the fault rocks reveal a syn-tectonic retrograde metamorphic evolution. Clasts of dolerite within the fault zone suggest that detachment faulting was accompanied by magmatic activity. Hydrothermal alteration is indicated by strong mineralogical and chemical modifications. Gabbro and serpentinized peridotite, together with serpentinite cataclasites occur as clasts in tectono-sedimentary breccias overlying directly the detachment fault. Across the whole Chenaillet Ophiolite, volcanic rocks directly overlie either the detachment fault or the sediments. In several places, N-S trending high-angle normal faults have been mapped. These faults truncate and displace the detachment fault leading to small domino-like structures. The basins, limited by these high-angle faults, are some hundreds to a few kilometres wide and few tens to some hundreds of meters deep. Because these high- angle faults are sealed locally by basalts and obliterated by volcanic structures, we interpret them as oceanic structures being active during the emplacement of the basalts. The alignment of porphyritic basaltic dykes parallel to, and their increasing abundance towards the high-angle faults suggest that they may have served as feeder channels for the overlying volcanic rocks. The complex poly-phase tectonic and magmatic processes observed in the Chenaillet Ophiolite are reminiscent of those reported from slow to ultraslow spreading ridges. The key result from our study is that mantle exhumation along detachment faults is followed by syn-magmatic normal faulting resulting in the emplacement of laterally variable, up to 300 meters thick massive lavas and pillow basalts covering the exhumed detachment fault. This implies that off-axis processes are more important as previously assumed and that large-scale detachment faults may be buried under massive volcanic sequences suggesting that detachment faulting is presumably more common than suggested by dredging or morpho-structural investigations of ultra- to slow- spreading oceanic crust.

The Death Valley turtlebacks reinterpreted as Miocene­ Pliocene folds of a major detachment surface

USGS Publications Warehouse

Holm, Daniel K.; Fleck, Robert J.; Lux, Daniel R.

1994-01-01

Determining the origin of extension parallel folds in metamorphic core complexes is fundamental to understanding the development of detachment faults. An excellent example of such a feature occurs in the Death Valley region of California where a major, undulatory, detachment fault is exposed along the well-known turtleback (antiformal) surfaces of the Black Mountains. In the hanging wall of this detachment fault are deformed strata of the Copper Canyon Formation. New age constraints indicate that the Copper Canyon Formation was deposited from ~6 to 3 Ma. The formation was folded during deposition into a SE-plunging syncline with an axial surface coplanar with that of a synform in the underlying detachment. This relation suggests the turtlebacks are a folded detachment surface formed during large-scale extension in an overall constrictional strain field. The present, more planar, Black Mountains frontal fault system may be the result of out-stepping of a normal fault system away from an older detachment fault that was deactivated by folding.

Chromosome Bridges Maintain Kinetochore-Microtubule Attachment throughout Mitosis and Rarely Break during Anaphase.

PubMed

Pampalona, Judit; Roscioli, Emanuele; Silkworth, William T; Bowden, Brent; Genescà, Anna; Tusell, Laura; Cimini, Daniela

2016-01-01

Accurate chromosome segregation during cell division is essential to maintain genome stability, and chromosome segregation errors are causally linked to genetic disorders and cancer. An anaphase chromosome bridge is a particular chromosome segregation error observed in cells that enter mitosis with fused chromosomes/sister chromatids. The widely accepted Breakage/Fusion/Bridge cycle model proposes that anaphase chromosome bridges break during mitosis to generate chromosome ends that will fuse during the following cell cycle, thus forming new bridges that will break, and so on. However, various studies have also shown a link between chromosome bridges and aneuploidy and/or polyploidy. In this study, we investigated the behavior and properties of chromosome bridges during mitosis, with the idea to gain insight into the potential mechanism underlying chromosome bridge-induced aneuploidy. We find that only a small number of chromosome bridges break during anaphase, whereas the rest persist through mitosis into the subsequent cell cycle. We also find that the microtubule bundles (k-fibers) bound to bridge kinetochores are not prone to breakage/detachment, thus supporting the conclusion that k-fiber detachment is not the cause of chromosome bridge-induced aneuploidy. Instead, our data suggest that while the microtubules bound to the kinetochores of normally segregating chromosomes shorten substantially during anaphase, the k-fibers bound to bridge kinetochores shorten only slightly, and may even lengthen, during anaphase. This causes some of the bridge kinetochores/chromosomes to lag behind in a position that is proximal to the cell/spindle equator and may cause the bridged chromosomes to be segregated into the same daughter nucleus or to form a micronucleus.

Pemphigus autoimmunity: Hypotheses and realities

PubMed Central

Grando, Sergei A

2011-01-01

The goal of contemporary research in pemphigus vulgaris and pemphigus foliaceus is to achieve and maintain clinical remission without corticosteroids. Recent advances of knowledge on pemphigus autoimmunity scrutinize old dogmas, resolve controversies, and open novel perspectives for treatment. Elucidation of intimate mechanisms of keratinocyte detachment and death in pemphigus has challenged the monopathogenic explanation of disease immunopathology. Over 50 organ-specific and non-organ-specific antigens can be targeted by pemphigus autoimmunity, including desmosomal cadherins and other adhesion molecules, PERP cholinergic and other cell membrane (CM) receptors, and mitochondrial proteins. The initial insult is sustained by the autoantibodies to the cell membrane receptor antigens triggering the intracellular signaling by Src, epidermal growth factor receptor kinase, protein kinases A and C, phospholipase C, mTOR, p38 MAPK, JNK, other tyrosine kinases, and calmodulin that cause basal cell shrinkage and ripping desmosomes off the CM. Autoantibodies synergize with effectors of apoptotic and oncotic pathways, serine proteases, and inflammatory cytokines to overcome the natural resistance and activate the cell death program in keratinocytes. The process of keratinocyte shrinkage/detachment and death via apoptosis/oncosis has been termed apoptolysis to emphasize that it is triggered by the same signal effectors and mediated by the same cell death enzymes. The natural course of pemphigus has improved due to a substantial progress in developing of the steroid-sparing therapies combining the immunosuppressive and direct anti-acantholytic effects. Further elucidation of the molecular mechanisms mediating immune dysregulation and apoptolysis in pemphigus should improve our understanding of disease pathogenesis and facilitate development of steroid-free treatment of patients. PMID:21939410

Hydrogels with Modulated Ionic Load for Mammalian Cell Harvesting with Reduced Bacterial Adhesion.

PubMed

Gallardo, Alberto; Martínez-Campos, Enrique; García, Carolina; Cortajarena, Aitziber L; Rodríguez-Hernández, Juan

2017-05-08

In this manuscript, we describe the fabrication of hydrogel supports for mammalian cell handling that can simultaneously prevent materials from microbial contamination and therefore allow storage in aqueous media. For that purpose, hydrogels based on the antifouling polymer polyvinylpyrrolidone (PVP) were functionalized with different ionic groups (anionic, cationic, or two types of zwitterions). In order to prevent bacterial adhesion in the long-term, we took advantage of the synergistic effect of inherently antifouling PVP and additional antifouling moieties incorporated within the hydrogel structure. We evaluated, in a separated series of experiments, both the capability of the materials to act as supports for the growth of mammalian cell monolayers for transplantation (using C-166-GFP endothelial cell line), as well their antifouling properties against Staphylococcus aureus, were studied. All of the hydrogels are structurally pseudodouble networks with high swelling (around 90%) and similar mechanical properties (in the low range for hydrogel materials with Young modulus below 1250 kPa). With some differences, all the charged hydrogels were capable of hosting mouse endothelial cell line C166-GFP to confluence, as well as a monolayer detachment and transplantation through simple mechanical agitation. On the contrary, the uncharged hydrogel was not capable to detach a full monolayer for transplantation. Bacterial adhesion and proliferation was highly sensitive to the functionality (type of charge and density). In particular, we evidenced that monomers bearing zwitterionic sulfobetaine groups, those negatively charged as well as "electro neutral" hydrogels fabricated from stoichiometric amounts of positive and negative units, exhibit excellent antifouling properties both at initial adhesion times and during longer periods up to 72 h.

Scar-free wound healing and regeneration following tail loss in the leopard gecko, Eublepharis macularius.

PubMed

Delorme, Stephanie Lynn; Lungu, Ilinca Mihaela; Vickaryous, Matthew Kenneth

2012-10-01

Many lizards are able to undergo scar-free wound healing and regeneration following loss of the tail. In most instances, lizard tail loss is facilitated by autotomy, an evolved mechanism that permits the tail to be self-detached at pre-existing fracture planes. However, it has also been reported that the tail can regenerate following surgical amputation outside the fracture plane. In this study, we used the leopard gecko, Eublepharis macularius, to investigate and compare wound healing and regeneration following autotomy at a fracture plane and amputation outside the fracture plane. Both forms of tail loss undergo a nearly identical sequence of events leading to scar-free wound healing and regeneration. Early wound healing is characterized by transient myofibroblasts and the formation of a highly proliferative wound epithelium immunoreactive for the wound keratin marker WE6. The new tail forms from what is commonly referred to as a blastema, a mass of proliferating mesenchymal-like cells. Blastema cells express the protease matrix metalloproteinase-9. Apoptosis (demonstrated by activated caspase 3 immunostaining) is largely restricted to isolated cells of the original and regenerating tail tissues, although cell death also occurs within dermal structures at the original-regenerated tissue interface and among clusters of newly formed myocytes. Furthermore, the autotomized tail is unique in demonstrating apoptosis among cells adjacent to the fracture planes. Unlike mammals, transforming growth factor-β3 is not involved in wound healing. We demonstrate that scar-free wound healing and regeneration are intrinsic properties of the tail, unrelated to the location or mode of tail detachment. Copyright © 2012 Wiley Periodicals, Inc.

Electronegative plasma diagnostic by laser photo-detachment combined with negatively biased Langmuir probe

NASA Astrophysics Data System (ADS)

Oudini, N.; Sirse, N.; Taccogna, F.; Ellingboe, A. R.; Bendib, A.

2018-05-01

We propose a new technique for diagnosing negative ion properties using Langmuir probe assisted pulsed laser photo-detachment. While the classical technique uses a laser pulse to convert negative ions into electron-atom pairs and a positively biased Langmuir probe tracking the change of electron saturation current, the proposed method uses a negatively biased Langmuir probe to track the temporal evolution of positive ion current. The negative bias aims to avoid the parasitic electron current inherent to probe tip surface ablation. In this work, we show through analytical and numerical approaches that, by knowing electron temperature and performing photo-detachment at two different laser wavelengths, it is possible to deduce plasma electronegativity (ratio of negative ion to electron densities) α, and anisothermicity (ratio of electron to negative ion temperatures) γ-. We present an analytical model that links the change in the collected positive ion current to plasma electronegativity and anisothermicity. Particle-In-Cell simulation is used as a numerical experiment covering a wide range of α and γ- to test the new analysis technique. The new technique is sensitive to α in the range 0.5 < α < 10 and yields γ- for large α, where negative ion flux affects the probe sheath behavior, typically α > 1.

Role of a reducing environment in disassembly of the herpesvirus tegument

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

Newcomb, William W.; Jones, Lisa M.; Dee, Alexander

2012-09-15

Initiation of infection by herpes family viruses involves a step in which most of the virus tegument becomes detached from the capsid. Detachment takes place in the host cell cytosol near the virus entry site and it is followed by dispersal of tegument proteins and disappearance of the tegument as a distinct entity. Here we describe the results of experiments designed to test the idea that the reducing environment of the cytosol may contribute to tegument detachment and disassembly. Non-ionic detergent was used to remove the membrane of purified herpes simplex virus under control and reducing conditions. The effects onmore » the tegument were then examined by SDS-PAGE and electron microscopy. Protein analysis demonstrated that most major tegument proteins were removed under both oxidizing and reducing conditions except for UL49 which required a reducing environment. It is proposed therefore that the reducing conditions in the cytosol are involved in removal of UL49 protein. Electron microscopic analysis revealed that capsids produced under oxidizing conditions contained a coating of protein that was absent in reduced virions and which correlated uniquely with the presence of UL49. This capsid-associated layer is suggested to be the location of UL49 in the extracted virion.« less

Mechanisms Underlying the Anti-Tumoral Effects of Citrus bergamia Juice

PubMed Central

Delle Monache, Simona; Sanità, Patrizia; Trapasso, Elena; Ursino, Maria Rita; Dugo, Paola; Russo, Marina; Ferlazzo, Nadia; Calapai, Gioacchino; Angelucci, Adriano; Navarra, Michele

2013-01-01

Based on the growing deal of data concerning the biological activity of flavonoid-rich natural products, the aim of the present study was to explore in vitro the potential anti-tumoral activity of Citrus Bergamia (bergamot) juice (BJ), determining its molecular interaction with cancer cells. Here we show that BJ reduced growth rate of different cancer cell lines, with the maximal growth inhibition observed in neuroblastoma cells (SH-SY5Y) after 72 hs of exposure to 5% BJ. The SH-SY5Y antiproliferative effect elicited by BJ was not due to a cytotoxic action and it did not induce apoptosis. Instead, BJ stimulated the arrest in the G1 phase of cell cycle and determined a modification in cellular morphology, causing a marked increase of detached cells. The inhibition of adhesive capacity on different physiologic substrates and on endothelial cells monolayer were correlated with an impairment of actin filaments, a reduction in the expression of the active form of focal adhesion kinase (FAK) that in turn caused inhibition of cell migration. In parallel, BJ seemed to hinder the association between the neural cell adhesion molecule (NCAM) and FAK. Our data suggest a mechanisms through which BJ can inhibit important molecular pathways related to cancer-associated aggressive phenotype and offer new suggestions for further studies on the role of BJ in cancer treatment. PMID:23613861

Photovoltaic solar concentrator

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

Nielson, Gregory N.; Cruz-Campa, Jose Luis; Okandan, Murat

A process including forming a photovoltaic solar cell on a substrate, the photovoltaic solar cell comprising an anchor positioned between the photovoltaic solar cell and the substrate to suspend the photovoltaic solar cell from the substrate. A surface of the photovoltaic solar cell opposite the substrate is attached to a receiving substrate. The receiving substrate may be bonded to the photovoltaic solar cell using an adhesive force or a metal connecting member. The photovoltaic solar cell is then detached from the substrate by lifting the receiving substrate having the photovoltaic solar cell attached thereto and severing the anchor connecting themore » photovoltaic solar cell to the substrate. Depending upon the type of receiving substrate used, the photovoltaic solar cell may be removed from the receiving substrate or remain on the receiving substrate for use in the final product.« less

Substance P prevents development of proliferative vitreoretinopathy in mice by modulating TNF-α

PubMed Central

Yoo, Kyungsang; Son, Bo Kwon; Kim, Suna; Son, Youngsook; Yu, Seung-Young

2017-01-01

Purpose Proliferative vitreoretinopathy (PVR) is an inflammatory fibrotic disease resulting from the inflammatory milieu after retinal detachment, which can prevent retinal healing. This study aimed to elucidate the effect of substance P (SP) on retinal degeneration caused by retinal detachment in vivo and to examine the role of SP in the tumor necrosis factor-alpha (TNF-α)-induced epithelial-mesenchymal transition (EMT) of human RPE cells in vitro. Methods PVR-like retinal damage was induced by intravitreally injecting dispase into mice, and SP was systemically injected twice a week for 3 weeks. Histological analysis and cytokine profile with enzyme-linked immunosorbent assay (ELISA) were performed. The direct effect of SP on induction of EMT in vitro was studied by adding SP to TNF-α-treated ARPE-19 cells and then evaluating the change in the characteristics of the epithelial and mesenchymal cells. Results Dispase injection led to a PVR-like retinal condition, demonstrating an inflammatory response with disruption of RPE interaction within 1 week and severe destruction with enfolding within 3 weeks after the dispase injection. The inflammatory environment promoted apoptosis and migration of fibroblast-like cells in the retinal layer, which can cause fibrotic disease, such as PVR. However, SP treatment suppressed early inflammatory responses by reducing TNF-α and elevating interleukin-10 (IL-10), with cell death and the appearance of fibroblastic cells inhibited and the progression of retinal degeneration obviously delayed. Moreover, SP ameliorated TNF-α-induced EMT of the RPE and directly prevented fibrotic change in the RPE. Conclusions This study revealed that SP can block apoptosis and EMT due to retinal inflammation and inhibit the development of PVR. This effect most likely occurred by modulating the secretion and action of TNF-α.. PMID:29296073

Requirements for cell rounding and surface protein down-regulation by Ebola virus glycoprotein.

PubMed

Francica, Joseph R; Matukonis, Meghan K; Bates, Paul

2009-01-20

Ebola virus causes an acute hemorrhagic fever that is associated with high morbidity and mortality. The viral glycoprotein is thought to contribute to pathogenesis, though precise mechanisms are unknown. Cellular pathogenesis can be modeled in vitro by expression of the Ebola viral glycoprotein (GP) in cells, which causes dramatic morphological changes, including cell rounding and surface protein down-regulation. These effects are known to be dependent on the presence of a highly glycosylated region of the glycoprotein, the mucin domain. Here we show that the mucin domain from the highly pathogenic Zaire subtype of Ebola virus is sufficient to cause characteristic cytopathology when expressed in the context of a foreign glycoprotein. Similarly to full length Ebola GP, expression of the mucin domain causes rounding, detachment from the extracellular matrix, and the down-regulation of cell surface levels of beta1 integrin and major histocompatibility complex class 1. These effects were not seen when the mucin domain was expressed in the context of a glycophosphatidylinositol-anchored isoform of the foreign glycoprotein. In contrast to earlier analysis of full length Ebola glycoproteins, chimeras carrying the mucin domains from the Zaire and Reston strains appear to cause similar levels of down-modulation and cell detachment. Cytopathology associated with Ebola glycoprotein expression does not occur when GP expression is restricted to the endoplasmic reticulum. In contrast to a previously published report, our results demonstrate that GP-induced surface protein down-regulation is not mediated through a dynamin-dependent pathway. Overall, these results support a model in which the mucin domain of Ebola GP acts at the cell surface to induce protein down modulation and cytopathic effects.

Rear Detachments: Capturing and Resourcing How the Army Fights

DTIC Science & Technology

2012-03-22

TDA ) for rear detachments to improve collective training and mission execution. 15. SUBJECT TERMS Rear Detachments, Doctrine, Organization, 16...as well as recommending that the Army establish a small Tables of Distribution and Allowances ( TDA ) for rear detachments to improve collective...and allowances ( TDA ), does not have substantive doctrine to guide its formation and operation, and generally organizes late in the ARFORGEN cycle

Light Scattering by Marine Particles: Modeling with Non-Spherical Shapes

DTIC Science & Technology

2011-04-15

scattering by nonspherical particles: application to coccoliths detached from Emiliania huxleyi. Limnology and Oceanography, 46. 1438— 1454,2001. H.R...application to coccoliths detached from Emiliania huxleyi," Limnol. Oceanogr. 46, 1438-1454 (2001). 5. H.R.Gordon, "Backscattering of light from...by coccoliths detached from Emiliania huxleyi," Applied Optics, 48, 6059-6073 (2009). Light scattering by coccoliths detached from Emiliania

Improvement of ore recovery efficiency in a flotation column cell using ultra-sonic enhanced bubbles

NASA Astrophysics Data System (ADS)

Filippov, L. O.; Royer, J. J.; Filippova, I. V.

2017-07-01

The ore process flotation technique is enhanced by using external ultra-sonic waves. Compared to the classical flotation method, the application of ultrasounds to flotation fluids generates micro-bubbles by hydrodynamic cavitation. Flotation performances increase was modelled as a result of increased probabilities of the particle-bubble attachment and reduced detachment probability under sonication. A simplified analytical Navier-Stokes model is used to predict the effect of ultrasonic waves on bubble behavior. If the theory is verified by experimentation, it predicts that the ultrasonic waves would create cavitation micro-bubbles, smaller than the flotation bubble added by the gas sparger. This effect leads to increasing the number of small bubbles in the liquid which promote particle-bubble attachment through coalescence between bubbles and micro-bubbles. The decrease in the radius of the flotation bubbles under external vibration forces has an additional effect by enhancing the bubble-particle collision. Preliminary results performed on a potash ore seem to confirm the theory.

Back to the Thymus: Peripheral T Cells Come Home

PubMed Central

Hale, J. Scott; Fink, Pamela J.

2009-01-01

The thymus has long been known as the generative organ for the T cell arm of the immune system. To perform this role, the thymus was thought to require protection from antigenic and cellular insult from the “outside world”, with the notable exception of the continual influx of progenitor cells required to initiate the complicated process of T cell differentiation. Overwhelming evidence that mature T cells can recirculate and persist in the thymus has required us to revamp this earlier view of the thymus as detached from outside influence. In this review, we consider the evidence for T cell recirculation into the thymus, discuss the likely means and location of mature T cell entry, and speculate on the potential consequences of such close apposition between differentiating thymocytes and mature recirculating lymphocytes. PMID:19030016

Back to the thymus: peripheral T cells come home.

PubMed

Hale, J Scott; Fink, Pamela J

2009-01-01

The thymus has long been known as the generative organ for the T-cell arm of the immune system. To perform this role, the thymus was thought to require protection from antigenic and cellular insult from the 'outside world', with the notable exception of the continual influx of progenitor cells required to initiate the complicated process of T-cell differentiation. Overwhelming evidence that mature T cells can recirculate and persist in the thymus has required us to revamp this earlier view of the thymus as detached from outside influence. In this review, we consider the evidence for T-cell recirculation into the thymus, discuss the likely means and location of mature T-cell entry, and speculate on the potential consequences of such close apposition between differentiating thymocytes and mature recirculating lymphocytes.

Defect, Kinetics and Heat Transfer of CDTE Bridgman Growth without Wall Contact

NASA Technical Reports Server (NTRS)

Larson, D. J., Jr.; Zhang, H.

2003-01-01

A detached growth mechanism has been proposed, which is similar to that proposed by Duffar et al. and used to study the current detached growth system. From numerical results, we can conclude that detached growth will more likely appear if the growth and wetting angles are large and meniscus is flat. Detached thickness is dependent on growth angle, wetting angle, and gap width and shape of the fins. The model can also explain why the detached growth will not happen for metals in which the growth angle is almost zero. Since the growth angle of CdZnTe cannot be changed, to promote detached growth, the number density of the fins should be low and the wetting angle should be high. Also, a much smaller gap width of the fins should be used in the ground experiment and the detached gap width is much smaller. The shape of the fins has minor influence on detached growth. An integrated numerical model for detached solidification has been developed combining a global heat transfer sub-model and a wall contact sub-model. The global heat transfer sub-model accounts for heat and mass transfer in the multiphase system, convection in the melt, macro-segregation, and interface dynamics. The location and dynamics of the solidification interface are accurately tracked by a multizone adaptive grid generation scheme. The wall contact sub-model accounts for the meniscus dynamics at the three-phase boundary. Simulations have been performed for crystal growth in a conventional ampoule and a designed ampoule to understand the benefits of detached solidification and its impacts on crystalline structural quality, e.g., stoichiometry, macro-segregation, and stress. From simulation results, both the Grashof and Marangoni numbers will have significant effects on the shape of growth front, Zn concentration distribution, and radial segregation. The integrated model can be used in designing apparatus and determining the optimal geometry for detached solidification in space and on the ground.

Abscisic Acid Accumulation by Roots of Xanthium strumarium L. and Lycopersicon esculentum Mill. in Relation to Water Stress.

PubMed

Cornish, K; Zeevaart, J A

1985-11-01

Plants of Xanthium strumarium L. and Lycopersicon esculentum Mill. cv ;Rheinlands Ruhm' were grown in solution culture, and control and steam-girdled intact plants were stressed. Detached roots of both species were stressed to different extents in two ways: (a) either in warm air or, (b) in the osmoticum Aquacide III. The roots of both species produced and accumulated progressively more abscisic acid (ABA), the greater the stress inflicted by either method. ABA-glucose ester levels in Xanthium roots were not affected by water stress and were too low to be the source of the stress-induced ABA. The fact that ABA accumulated in detached roots and in roots of girdled plants proves that ABA was synthesized in the roots and not merely transported from the shoots.Maximum ABA accumulation in detached roots occurred after 60 to 70% loss of fresh weight. In Xanthium roots, ABA levels continued to increase for at least 11 hours, and no catabolism was apparent when stressed roots were immersed in water, although the roots did stop accumulating ABA. When osmotically stressed, Xanthium roots reached a maximum ABA level after 2 hours, but ABA continued to rise in the medium.Under optimal stress conditions, endogenous ABA levels increased 100 times over their prestress values in detached roots of Xanthium, and 15 times in Lycopersicon under nonoptimal stress, when endogenous ABA was expressed as concentrations based on tissue water content. These are much greater relative increases than observed in the leaves (15 times in Xanthium, 3 times in Lycopersicon), although the roots contain substantially less ABA than the leaves in all circumstances. The results suggest that the endogenous level of ABA in roots could rise appreciably prior to leaf wilt, and could modify the plant's water economy before the leaves become stressed.

Abscisic Acid Accumulation by Roots of Xanthium strumarium L. and Lycopersicon esculentum Mill. in Relation to Water Stress 1

PubMed Central

Cornish, Katrina; Zeevaart, Jan A. D.

1985-01-01

Plants of Xanthium strumarium L. and Lycopersicon esculentum Mill. cv `Rheinlands Ruhm' were grown in solution culture, and control and steam-girdled intact plants were stressed. Detached roots of both species were stressed to different extents in two ways: (a) either in warm air or, (b) in the osmoticum Aquacide III. The roots of both species produced and accumulated progressively more abscisic acid (ABA), the greater the stress inflicted by either method. ABA-glucose ester levels in Xanthium roots were not affected by water stress and were too low to be the source of the stress-induced ABA. The fact that ABA accumulated in detached roots and in roots of girdled plants proves that ABA was synthesized in the roots and not merely transported from the shoots. Maximum ABA accumulation in detached roots occurred after 60 to 70% loss of fresh weight. In Xanthium roots, ABA levels continued to increase for at least 11 hours, and no catabolism was apparent when stressed roots were immersed in water, although the roots did stop accumulating ABA. When osmotically stressed, Xanthium roots reached a maximum ABA level after 2 hours, but ABA continued to rise in the medium. Under optimal stress conditions, endogenous ABA levels increased 100 times over their prestress values in detached roots of Xanthium, and 15 times in Lycopersicon under nonoptimal stress, when endogenous ABA was expressed as concentrations based on tissue water content. These are much greater relative increases than observed in the leaves (15 times in Xanthium, 3 times in Lycopersicon), although the roots contain substantially less ABA than the leaves in all circumstances. The results suggest that the endogenous level of ABA in roots could rise appreciably prior to leaf wilt, and could modify the plant's water economy before the leaves become stressed. PMID:16664467

A Meta-Analysis on Antecedents and Outcomes of Detachment from Work.

PubMed

Wendsche, Johannes; Lohmann-Haislah, Andrea

2016-01-01

Detachment from work has been proposed as an important non-work experience helping employees to recover from work demands. This meta-analysis (86 publications, k = 91 independent study samples, N = 38,124 employees) examined core antecedents and outcomes of detachment in employee samples. With regard to outcomes, results indicated average positive correlations between detachment and self-reported mental (i.e., less exhaustion, higher life satisfaction, more well-being, better sleep) and physical (i.e., lower physical discomfort) health, state well-being (i.e., less fatigue, higher positive affect, more intensive state of recovery), and task performance (small to medium sized effects). However, average relationships between detachment and physiological stress indicators and work motivation were not significant while associations with contextual performance and creativity were significant, but negative. Concerning work characteristics, as expected, job demands were negatively related and job resources were positively related to detachment (small sized effects). Further, analyses revealed that person characteristics such as negative affectivity/neuroticism (small sized effect) and heavy work investment (medium sized effect) were negatively related to detachment whereas detachment and demographic variables (i.e., age and gender) were not related. Moreover, we found a medium sized average negative relationship between engagement in work-related activities during non-work time and detachment. For most of the examined relationships heterogeneity of effect sizes was moderate to high. We identified study design, samples' gender distribution, and affective valence of work-related thoughts as moderators for some of these aforementioned relationships. The results of this meta-analysis point to detachment as a non-work (recovery) experience that is influenced by work-related and personal characteristics which in turn is relevant for a range of employee outcomes.

A Meta-Analysis on Antecedents and Outcomes of Detachment from Work

PubMed Central

Wendsche, Johannes; Lohmann-Haislah, Andrea

2017-01-01

Detachment from work has been proposed as an important non-work experience helping employees to recover from work demands. This meta-analysis (86 publications, k = 91 independent study samples, N = 38,124 employees) examined core antecedents and outcomes of detachment in employee samples. With regard to outcomes, results indicated average positive correlations between detachment and self-reported mental (i.e., less exhaustion, higher life satisfaction, more well-being, better sleep) and physical (i.e., lower physical discomfort) health, state well-being (i.e., less fatigue, higher positive affect, more intensive state of recovery), and task performance (small to medium sized effects). However, average relationships between detachment and physiological stress indicators and work motivation were not significant while associations with contextual performance and creativity were significant, but negative. Concerning work characteristics, as expected, job demands were negatively related and job resources were positively related to detachment (small sized effects). Further, analyses revealed that person characteristics such as negative affectivity/neuroticism (small sized effect) and heavy work investment (medium sized effect) were negatively related to detachment whereas detachment and demographic variables (i.e., age and gender) were not related. Moreover, we found a medium sized average negative relationship between engagement in work-related activities during non-work time and detachment. For most of the examined relationships heterogeneity of effect sizes was moderate to high. We identified study design, samples' gender distribution, and affective valence of work-related thoughts as moderators for some of these aforementioned relationships. The results of this meta-analysis point to detachment as a non-work (recovery) experience that is influenced by work-related and personal characteristics which in turn is relevant for a range of employee outcomes. PMID:28133454

Effect of ellagic acid on proliferation, cell adhesion and apoptosis in SH-SY5Y human neuroblastoma cells.

PubMed

Fjaeraa, Christina; Nånberg, Eewa

2009-05-01

Ellagic acid, a polyphenolic compound found in berries, fruits and nuts, has been shown to possess growth-inhibiting and apoptosis promoting activities in cancer cell lines in vitro. The objective of this study was to investigate the effect of ellagic acid in human neuroblastoma SH-SY5Y cells. In cultures of SH-SY5Y cells incubated with ellagic acid, time- and concentration-dependent inhibitory effects on cell number were demonstrated. Ellagic acid induced cell detachment, decreased cell viability and induced apoptosis as measured by DNA strand breaks. Ellagic acid-induced alterations in cell cycle were also observed. Simultaneous treatment with all-trans retinoic acid did not rescue the cells from ellagic acid effects. Furthermore, the results suggested that pre-treatment with all-trans retinoic acid to induce differentiation and cell cycle arrest did not rescue the cells from ellagic acid-induced cell death.

Sensitivity of small myosin II ensembles from different isoforms to mechanical load and ATP concentration.

PubMed

Erdmann, Thorsten; Bartelheimer, Kathrin; Schwarz, Ulrich S

2016-11-01

Based on a detailed crossbridge model for individual myosin II motors, we systematically study the influence of mechanical load and adenosine triphosphate (ATP) concentration on small myosin II ensembles made from different isoforms. For skeletal and smooth muscle myosin II, which are often used in actomyosin gels that reconstitute cell contractility, fast forward movement is restricted to a small region of phase space with low mechanical load and high ATP concentration, which is also characterized by frequent ensemble detachment. At high load, these ensembles are stalled or move backwards, but forward motion can be restored by decreasing ATP concentration. In contrast, small ensembles of nonmuscle myosin II isoforms, which are found in the cytoskeleton of nonmuscle cells, are hardly affected by ATP concentration due to the slow kinetics of the bound states. For all isoforms, the thermodynamic efficiency of ensemble movement increases with decreasing ATP concentration, but this effect is weaker for the nonmuscle myosin II isoforms.

Free-standing polyelectrolyte membranes made of chitosan and alginate

PubMed Central

Caridade, Sofia G.; Monge, Claire; Gilde, Flora; Boudou, Thomas; Mano, João F.; Picart, Catherine

2014-01-01

Free-standing films have increasing applications in the biomedical field as drug delivery systems, for wound healing and tissue engineering. Here, we prepared free-standing membranes by the layer-by-layer assembly of chitosan and alginate, two widely used biomaterials. Our aim was to produce thick membrane, to study the permeation of model drugs and the adhesion of muscle cells. We first defined the optimal growth conditions in terms of pH and alginate concentration. The membranes could be easily detached from polystyrene or polypropylene substrate without any post-processing step. They dry thickness was varied over a large range from 4 to 35 μm. A two-fold swelling was observed by confocal microscopy when they were immersed in PBS. In addition, we quantified the permeation of model drugs (fluorescent dextrans) through the free standing membrane, which depended on the dextran molecular weight. Finally, we showed that myoblast cells exhibited a preferential adhesion on the alginate-ending membrane as compared to the chitosan-ending membrane or to the substrate side. PMID:23590116

Antitumor Effects In Vitro and In Vivo and Mechanisms of Protection against Melanoma B16F10-Nex2 Cells By Fastuosain, a Cysteine Proteinase from Bromelia fastuosa1

PubMed Central

Guimarães-Ferreira, Carla A; Rodrigues, Elaine G; Mortara, Renato A; Cabral, Hamilton; Serrano, Fabiana A; Ribeiro-dos-Santos, Ricardo; Travassos, Luiz R

2007-01-01

In the present work, the antitumor effect of fastuosain, a cysteine proteinase from Bromelia fastuosa, was investigated. In the intravenous model of lung colonization in C57Bl/6 mice, fastuosain and bromelain injected intraperitoneally were protective, and very few nodules of B16F10-Nex2 melanoma cells were detected. Tumor cells treated with fastuosain showed reduced expression of CD44 and decreased invasion through Matrigel, lost their cytoplasmic extensions and substrate adherence, and became round and detached, forming strongly bound cell clusters in suspension. Peritoneal cells recruited and activated by fastuosain treatment (mainly monocytic cells and lymphocytes) migrated to the lung, where pulmonary melanoma metastases grew. Adoptive transference of peritoneal cells recruited by fastuosain had no protective effect against lung metastases in recipient mice. Treatment of green fluorescent protein-chimeric animals with fastuosain did not change the number of cells that migrated to the lung, compared to PBS-injected control mice, but the number of positive major histocompatibility complex class II cells increased with fastuosain treatment. Murine antibodies against fastuosain, bromelain, and cathepsins B and L cross-reacted in ELISA and recognized surface and cytoplasmic components expressed on B16F10-Nex2 cells. Anti-fastuosain antibodies were cytotoxic/lytic to B16F10-Nex2 cells. Antitumor effects of fastuosain involve mainly the direct effect of the enzyme and elicitation of protective antibodies. PMID:17898868

Effect of fuel concentration and force on collective transport by a team of dynein motors

PubMed Central

Takshak, Anjneya; Roy, Tanushree; Tandaiya, Parag

2016-01-01

Abstract Motor proteins are essential components of intracellular transport inside eukaryotic cells. These protein molecules use chemical energy obtained from hydrolysis of ATP to produce mechanical forces required for transporting cargos inside cells, from one location to another, in a directed manner. Of these motors, cytoplasmic dynein is structurally more complex than other motor proteins involved in intracellular transport, as it shows force and fuel (ATP) concentration dependent step‐size. Cytoplasmic dynein motors are known to work in a team during cargo transport and force generation. Here, we use a complete Monte‐Carlo model of single dynein constrained by in vitro experiments, which includes the effect of both force and ATP on stepping as well as detachment of motors under force. We then use our complete Monte‐Carlo model of single dynein motor to understand collective cargo transport by a team of dynein motors, such as dependence of cargo travel distance and velocity on applied force and fuel concentration. In our model, cargos pulled by a team of dynein motors do not detach rapidly under higher forces, confirming the experimental observation of longer persistence time of dynein team on microtubule under higher forces. PMID:27727483

The conserved transmembrane proteoglycan Perdido/Kon-tiki is essential for myofibrillogenesis and sarcomeric structure in Drosophila

PubMed Central

Pérez-Moreno, Juan J.; Bischoff, Marcus; Martín-Bermudo, Maria D.; Estrada, Beatriz

2014-01-01

ABSTRACT Muscle differentiation requires the assembly of high-order structures called myofibrils, composed of sarcomeres. Even though the molecular organization of sarcomeres is well known, the mechanisms underlying myofibrillogenesis are poorly understood. It has been proposed that integrin-dependent adhesion nucleates myofibrils at the periphery of the muscle cell to sustain sarcomere assembly. Here, we report a role for the gene perdido (perd, also known as kon-tiki, a transmembrane chondroitin proteoglycan) in myofibrillogenesis. Expression of perd RNAi in muscles, prior to adult myogenesis, can induce misorientation and detachment of Drosophila adult abdominal muscles. In comparison to controls, perd-depleted muscles contain fewer myofibrils, which are localized at the cell periphery. These myofibrils are detached from each other and display a defective sarcomeric structure. Our results demonstrate that the extracellular matrix receptor Perd has a specific role in the assembly of myofibrils and in sarcomeric organization. We suggest that Perd acts downstream or in parallel to integrins to enable the connection of nascent myofibrils to the Z-bands. Our work identifies the Drosophila adult abdominal muscles as a model to investigate in vivo the mechanisms behind myofibrillogenesis. PMID:24794494

Candida tropicalis biofilms: artificial urine, urinary catheters and flow model.

PubMed

Negri, Melyssa; Silva, Sónia; Henriques, Mariana; Azeredo, Joana; Svidzinski, Terezinha; Oliveira, Rosário

2011-10-01

Adhesion to medical devices and biofilm formation are considered important virulence factors of Candida tropicalis. This work aimed to use artificial urine (AU) and urinary catheters, under flow conditions, for studying C. tropicalis biofilms. Adhesion and biofilm formation on silicone and latex urinary catheters were quantified by crystal violet staining and determination of colony forming units. Candida surface hydrophobicity was also evaluated, as well as the biofilms' matrix content in terms of proteins and carbohydrates. Candida tropicalis was able to adhere and to form biofilms along the entire length of the catheters under flow conditions. It was found that the isolate U69 adhered significantly more to both types of catheters than did the reference strain. However, U69 biofilms contained significantly less cultivable cells and higher biofilm biomass than those of the reference strain. Detachment of cells from biofilms on latex catheter was lower compared to silicone catheter. This model using AU appeared to be suitable for studies mimicking the real body conditions. Additionally, C. tropicalis was in fact able to colonize urinary catheters in the presence of AU and to detach from these catheters, demonstrating their capacity to colonize distal sites.

Strategy for the management of uncomplicated retinal detachments: the European vitreo-retinal society retinal detachment study report 1.

PubMed

Adelman, Ron A; Parnes, Aaron J; Ducournau, Didier

2013-09-01

To study success and failure in the treatment of uncomplicated rhegmatogenous retinal detachments (RRDs). Nonrandomized, multicenter retrospective study. One hundred seventy-six surgeons from 48 countries spanning 5 continents provided information on the primary procedures for 7678 cases of RRDs including 4179 patients with uncomplicated RRDs. Reported data included specific clinical findings, the method of repair, and the outcome after intervention. Final failure of retinal detachment repair (level 1 failure rate), remaining silicone oil at the study's conclusion (level 2 failure rate), and need for additional procedures to repair the detachment (level 3 failure rate). Four thousand one hundred seventy-nine uncomplicated cases of RRD were included. Combining phakic, pseudophakic, and aphakic groups, those treated with scleral buckle alone (n = 1341) had a significantly lower final failure rate than those treated with vitrectomy, with or without a supplemental buckle (n = 2723; P = 0.04). In phakic patients, final failure rate was lower in the scleral buckle group compared with those who had vitrectomy, with or without a supplemental buckle (P = 0.028). In pseudophakic patients, the failure rate of the initial procedure was lower in the vitrectomy group compared with the scleral buckle group (P = 3×10(-8)). There was no statistically significant difference in failure rate between segmental (n = 721) and encircling (n = 351) buckles (P = 0.5). Those who underwent vitrectomy with a supplemental scleral buckle (n = 488) had an increased failure rate compared with those who underwent vitrectomy alone (n = 2235; P = 0.048). Pneumatic retinopexy was found to be comparable with scleral buckle when a retinal hole was present (P = 0.65), but not in cases with a flap tear (P = 0.034). In the treatment of uncomplicated phakic retinal detachments, repair using scleral buckle may be a good option. There was no significant difference between segmental versus 360-degree buckle. For pseudophakic uncomplicated retinal detachments, the surgeon should balance the risks and benefits of vitrectomy versus scleral buckle and keep in mind that the single-surgery reattachment rate may be higher with vitrectomy. However, if a vitrectomy is to be performed, these data suggest that a supplemental buckle is not helpful. The author(s) have no proprietary or commercial interest in any materials discussed in this article. Copyright © 2013 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.

Tractional retinal detachment in Usher syndrome type II.

PubMed

Rani, Alka; Pal, Nikhil; Azad, Raj Vardhan; Sharma, Yog Raj; Chandra, Parijat; Vikram Singh, Deependra

2005-08-01

Retinal detachment is a rare complication in patients with retinitis pigmentosa. A case is reported of tractional retinal detachment in a patient with retinitis pigmentosa and sensorineural hearing loss, which was diagnosed as Usher syndrome type II. Because of the poor visual prognosis, the patient refused surgery in that eye. Tractional retinal detachment should be added to the differential diagnoses of visual loss in patients with retinitis pigmentosa.

Adhesion Upon Solidification and Detachment in the Melt Spinning of Metals

NASA Astrophysics Data System (ADS)

Altieri, Anthony L.; Steen, Paul H.

2014-12-01

In planar-flow melt spinning, liquid metal is rapidly solidified, against a heat-sink wheel, into thin ribbons which adhere to the substrate wheel. In the absence of a blade to mechanically scrape the ribbon off the wheel, it may wrap fully around and re-enter the solidification region, called `catastrophic' adhesion. Otherwise, detachment occurs part way around the wheel, called `natural' detachment. Natural detachment occurs through a release of thermo-elastic stress after sufficient cooling of the ribbon, according to prior studies. This note extends prior work by invoking a crack propagation view of natural detachment which, when combined with a simple model of the thermo-elastic stress build-up and ribbon cooling, yields an adhesion/detachment criterion characterized by an interfacial adhesion/fracture energy . For aluminum-silicon alloys frozen against a copper substrate, we report 60 N/m. The criterion can be used to predict detachment once a heat-transfer coefficient is known. We obtain this parameter from natural detachment experiments and then use it to predict catastrophic adhesion in a semi-empirical way. Our note puts a quantitative foundation underneath prior qualitative discussions in the literature. Alternatively, it demonstrates how the interfacial strength of adhesion, a property only of the pair of adhering materials, might be measured based on sticking distance experiments.

Post-Mortem Pathological Examination of Two Patients after Intraaneurysmal Embolization Using Guglielmi Detachable Coils

PubMed Central

Nakahara, T.; Sakamoto, S.; Hamasaki, O.; Sakoda, K.

2003-01-01

Summary We report the histological findings in two patients treated using Guglielmi detachable coils with almost complete occlusion of the aneurysms. Autopsies of these patients were performed one week and one year after GDC embolization respectively. In one aneurysm that was obtained at autopsy one week after embolization, the histological findings revealed coils and an unorganized thrombus-filled aneurysm sac; an incomplete cell-lining on the luminal side of fibrin thrombi in the region of the neck of the aneurysm was recognized. In the other aneurysm in which autopsy was performed one year after embolization, an organized fibrous tissue at the margin of the aneurysmal wall and vascular granulation tissue at the center of the aneurysm were observed. There is a single layer of endothelium covering fibrous tissue in the neck of the aneurysm. We discuss the healing process after GDC treatment. PMID:20591231