Role of E-Cadherin in Membrane-Cortex Interaction Probed by Nanotube Extrusion

This study aims to define the role of E-cadherin (Ecad) engagement in cell-cell contact during membrane-cortex interaction. As a tool, we used a hydrodynamic membrane tube extrusion technique to characterize the mechanical interaction between the plasma membrane and the underlying cortical cytoskeleton. Cells were anchored on 4.5 μm beads coated with polylysine (PL) to obtain nonspecific cell adhesion or with an antibody against Ecad to mimic specific Ecad-mediated cell adhesion. We investigated tube length dynamics L(t) over time and through successive extrusions applied to the cell at regular time intervals. A constant slow velocity was observed for the first extrusion, for PL-attached cells. Subsequent extrusions had two phases: an initial high-velocity regime followed by a low-velocity regime. Successive extrusions gradually weakened the binding of the membrane around the tube neck to the underlying cortical cytoskeleton. Cells specifically attached via Ecad first exhibited a very low extrusion velocity regime followed by a faster extrusion regime similar to nonspecific extrusion. This indicates that Ecad strengthens the membrane-cortical cytoskeleton interaction, but only in a restricted area corresponding to the site of contact between the cell and the bead. Occasional giant “cortex” tubes were extruded with specifically anchored cells, demonstrating that the cortex remained tightly bound to the membrane through Ecad-mediated adhesion at the contact site.     

Patient Preferences Regarding Device Reuse and Potential of Devices for Reuse - A Study in a Veteran Population

BackgroundMany cardiac patients need and undergo device implants. Veterans’ preferences regarding post-mortem handling of devices are not known. Cardiac patients in low- and middle-income countries who need but cannot afford devices rely on donations. Charitable organizations have successfully provided devices for reuse to such patients.ObjectiveWe estimated the number of devices with potential for possible reuse in a veteran population.MethodsBetween January and December 2008, at a tertiary medical center, veterans with implanted cardiac devices were surveyed using a questionnaire for their preferences regarding post-mortem handling. One choice was donation to charity for reuse. Although altruistic, it is unclear what percent of such devices have reuse potential. Retrospective chart review of veterans who underwent device implants between 1992 and 2007 identified a cohort of patients with Implantable Cardiac Defibrillators (ICDs) who had died by April 31st 2009. In this cohort, ICDs implanted in the year preceding the patient’s death were counted as having reuse potential.Results94 of 97 veterans completed the survey. 56% were unaware of how devices are handled after death. The top three preferences for postmortem handling were: return to manufacturer, return to hospital and donation for reuse. 88% were willing to sign an advance device directive. Retrospective review identified 161 veterans who had received 301 ICDs. Of these, 77 ICDs (25%) had median reuse potential of 3.1 years.ConclusionIn a VA cohort of deceased patients a substantial proportion of devices had reuse potential. Further research is needed to direct health policy.     

Three-Dimensional Computational Modeling of an Extra-Descending Aortic Assist Device Using Fluid-Structure Interaction

BackgroundThe incidence of heart failure is anticipated to rise by 2030, resulting in more than 8 million adults with this condition in US. Despite the advancement in pharmacological and surgical treatments, some patients progress to severe forms of cardiac dysfunction requiring cardiac transplantation as a last-resort treatment. Cardiac assist devices play an essential role in the recovery of normal cardiac performance through reversible remodeling or in assisting the weak organ to prolong survival rate. However, these devices need to be monitored carefully, as prolonged use may lead to physiological maladaptation and further cardiac complications. The optimization of such devices has done through the development and use of numerical simulations that allow the analysis of in-vivo hemodynamic patterns of blood flow. This study aims to investigate the performance of a model of extra-aortic assist device surrounding the descending aorta through three-dimensional patient-specific modeling.MethodsA three-dimensional model of the aorta was constructed from patient-specific cardiac CT images of a 60-year-old male diagnosed with left ventricular failure at the Tehran Heart Center (THC). Numerical simulation was conducted for two complete cardiac cycles using fluid-structure interaction (FSI) analysis under the assumption that the balloon and the aortic vessel behave as linear elastic materials, and that blood is a Newtonian and incompressible fluid.ResultsThe numerical simulation demonstrated a high correlation between the FSI analysis and clinical data of the patient-specific anatomical and physiological conditions. Blood velocity, pressure, deformation, and strain contours were simulated and analyzed through three-dimensional modeling. Compared to the unassisted aorta, the device provided an increase in blood flow displacement of an additional 15 ml of blood in the descending aorta, brachiocephalic, carotid, and subclavian arteries. The maximum von Mises stress distribution across the aortic vessel was higher than the stress imposed on the system in the unassisted heart, with values of 3.3 MPa and 0.28 MPa, respectively. Numerical investigation of structural responses revealed that no remarkable force was exerted on the aortic valve by the device at the descending aorta.ConclusionWe present the numerical investigation of a counterpulsation device around the descending aorta that has not previously been tested on human or animal models. While this extra-aortic balloon pump (EABP) did not show a significant improvement in coronary perfusion, there is room for improvement in further studies to optimize the geometry of the balloon. Additional investigations are required to determine the efficacy of this device and its safety before in-vivo experimental studies are pursued. This simulation has clinical relevance when choosing an appropriate cardiac assist device to address patient-specific physiological and pathological conditions.     

Ultrasound-guided axillary vein puncture for cardiac devices implantation in patients under antithrombotic therapy

BackgroundUltrasound-guided axillary venous puncture (UGAVP) for cardiac devices implantation has been developed because of its rapidity, safety and potential long-term lead protection. Early work excluded defibrillators (ICD), cardiac resynchronization therapy (CRT) and upgrade procedures. Compared to the cephalic approach, in previous studies, there was a greater use of pressure dressings with this technique, suggesting a higher risk of bleeding.AimsTo assess UGAVP in patients under antithrombotic therapy (ATT) undergoing cardiac devices implantation including CRT/ICD.MethodsProspectively, consecutive patients eligible for a pacemaker or ICD implantation were included. All procedures were performed by a single operator, experienced with UGAVP for femoral access, and fluoroscopy-guided axillary vein access. Guidewires insertion time (from lidocaïne administration), and complications were systematically studied.ResultsFrom 457 cardiac device implantations, 200 patients (77.8 ± 10 y, male 58%) 360 leads were implanted by UGAVP including 36 ICD, 54 CRT and 14 upgrade procedures. A majority (90%) was under ATT: Vitamin K Antagonist or Heparin (n = 58, 29%), direct oral anticoagulant (n = 46, 23%), dual antithrombotic therapy (n = 18, 9%) and single antiplatelet drug (n = 82, 41%). UGAVP was successful in 95.78%. Mean insertion time for 1.8 guidewires per patient was 4.68 ± 3.6 min. No complication (no hematoma) was observed during the follow-up (mean of 45 ± 10 months). Guidewires insertion time reached its plateau after 15 patients.ConclusionUGAVP is fast, feasible and safe for patients under ATT undergoing device implantation including CRT/ICD and upgrade procedures, with a short learning curve.     

Early recurrence of thoracolumbar intervertebral disc extrusion after surgical decompression: a report of three cases

Thoracolumbar disc extrusions were diagnosed in three chondrodystrophic dogs with paraparesis of up to three days duration. All cases were managed by hemilaminectomy and removal of extruded disc material. In one dog, fenestration of the herniated disc space was also performed. Initially neurological function improved or was unchanged, but from two to ten days postoperatively clinical signs of deterioration became apparent. In all the dogs, recurrence of disc extrusion at the same location as the initial extrusion was diagnosed by computer tomography and at a second surgery abundant disc material was found at the hemilaminectomy site between the dura and an implanted graft of autogenous fat.     

The fate of breast implants with infections around them.

In a retrospective study of 41 infected breasts following the insertion of implants, a high incidence of postoperative hematoma was noted. When infection occurred, cultures usually demonstrated the causative organism to be Staphylococcus aureus. Treatment by conservative drainage and vigorous antibiotics was generally successful in salvaging those implants which had been inserted through an areolar incision for simple augmentation, or under the pectoralis muscle (or a dermal pedicle) after a subcutaneous mastectomy. Those patients whose augmentations had been done through an inframammary incision, or whose implant after a subcutaneous mastectomy was under the skin flap, had a statistically higher incidence of implant loss. Most breasts will salvaged implants became firm.     

A Mechanical Instability in Planar Epithelial Monolayers Leads to Cell Extrusion

In cell extrusion, a cell embedded in an epithelial monolayer loses its apical or basal surface and is subsequently squeezed out of the monolayer by neighboring cells. Cell extrusions occur during apoptosis, epithelial-mesenchymal transition, or precancerous cell invasion. They play important roles in embryogenesis, homeostasis, carcinogenesis, and many other biological processes. Although many of the molecular factors involved in cell extrusion are known, little is known about the mechanical basis of cell extrusion. We used a three-dimensional (3D) vertex model to investigate the mechanical stability of cells arranged in a monolayer with 3D foam geometry. We found that when the cells composing the monolayer have homogeneous mechanical properties, cells are extruded from the monolayer when the symmetry of the 3D geometry is broken because of an increase in cell density or a decrease in the number of topological neighbors around single cells. Those results suggest that mechanical instability inherent in the 3D foam geometry of epithelial monolayers is sufficient to drive epithelial cell extrusion. In the situation in which cells in the monolayer actively generate contractile or adhesive forces under the control of intrinsic genetic programs, the forces act to break the symmetry of the monolayer, leading to cell extrusion that is directed to the apical or basal side of the monolayer by the balance of contractile and adhesive forces on the apical and basal sides. Although our analyses are based on a simple mechanical model, our results are in accordance with observations of epithelial monolayers in vivo and consistently explain cell extrusions under a wide range of physiological and pathophysiological conditions. Our results illustrate the importance of a mechanical understanding of cell extrusion and provide a basis by which to link molecular regulation to physical processes.     

Time course of meiotic progression after transferring primary spermatocyte into ooplasm at different stages

This study attempted to investigate the time course of meiotic progression after transferring primary spermatocyte (PS) into ooplasm at different maturing stages. In present experiments, PSs were introduced into maturing ooplasts or oocytes by electrofusion. Higher fusion rate was obtained by phytohemagglutinin (PHA) agglutination than by perivitelline space (PVS) insertion. When the ooplasms prepared at 0, 2, 5, and 8.5 hr of in vitro maturation (IVM) were used as recipients and PSs were used as donors, the reconstructed cells extruded the first polar body (PB1) approximately 8.5, 7, 5.5, and 3 hr after electrofusion, respectively. Especially, when ooplasm cultured for 8.5 hr in vitro after GV removal was fused with PS, the PB1 was emitted 7-11 hr after electrofusion. Additionally, the PB1 extrusions of GV and pro-MI oocytes fertilized with PSs were 2.5 hr earlier than control oocytes. The results suggest that (1) PSs undergo the first meiosis in different time courses when introduced into ooplasm at different maturing stages; (2) GV material plays an important role in determining the timing of PB1 extrusion; and (3) first meiotic division of GV and pro-MI oocytes can be accelerated by introducing PS.     

Calling behaviour in arctiid moths: The effects of temperature and wind speed on the rhythmic exposure of the sex attractant gland

Female arctiid moths rhythmically extrude their ovipositors and thereby rhythmically expose their sex pheromone glands while calling. In Utetheisa ornatrix this behaviour results in concentration-modulated or pulsed pheromone release. The ovipositor protrusion frequencies were determined for 15 arctiid species: U. ornatrix, Haploa clymene, H. colona, H. lecontei, Pyrrarctia isabella, Estigmene acrea, Spilosoma congrua, S. virginica, Ecpantheria scribonia, Phragmatobia fuliginosa, Apantesis nais, A. arge, Pareuchaetes insulata, Cycnia tenera, and Euchaetes egle. Moth species that call early in the scotophase extrude their ovipositors at high frequencies (up to 170.9 ± 24.9 extrusions/min at 25°C) while moth species that call late in scotophase extrude their ovipositors at low frequencies (down to 68.3 ± 5.4 extrusions/min at 25°C). In all species tested, the ovipositor extrusion frequencies were shown to be temperature sensitive with a mean Q₁₀ of 2.0 ± 0.1. In one species, U. ornatrix, the ovipositor extrusion frequency varies with wind speed. In still air the ovipositor extrusion frequency is 70.1 ± 8.4 extrusions/min and at a wind speed of 120 cm/s the ovipositor extrusion frequency is 142.1 ± 8.3 extrusions/min. Suggested functions for rhythmic ovipositor extrusion during calling are discussed.     

Evaluation of a New Extrusion Device for the Production of Stable Oligolamellar Liposomes in a Liter Scale

Abstract

A procedure for large-scale production of extruded oligolamellar liposomes was developed. the extrusion technique described is rapid, simple, and reproducible. Vesicles with a diameter in the range of 200-500 nm are obtained. the liposomal preparations were characterized by their specific turbidity (determined spectrophotometrically) and contamination with large particles (> 1 µm). Liposome physical stability was determined using a parameter (P) that relates the ratio of the specific turbidity before and after standardized centrifugation.

The extruded oligolamellar liposomes obtained were stable, and after 4 weeks storage at 4°C no irreversible aggregation or fusion of the vesicles occurs.

The extrusion process ensures sterilization of the final liposomal product, making it acceptable as a pharmaceutical preparation for parenteral use.     

Optim-Home : optimisation de la prise en charge de patients insuffisants cardiaques de l’hôpital à leur domicile par home monitoring

The Optim Home project concerns cardiac heart failure recipients subjected to cardiac resynchronization therapy. Its objective is the development of optimal cardiac resynchronization devices. Several questions were addressed: i) definition of a new tool for optimal positioning of embedded or external sensors; ii) development of embedded algorithms to improve the delivery of the therapy; iii) proposition of data mining algorithms for remote monitoring of heart failure patients; iv) conception of an “home-monitoring” platform for data processing and secure data-transfer from an implanted device and health monitoring center. At the end, the Optim Home project will lead to the conception of a communicating device optimizing the management of heart failure patients and their long-term monitoring at home.     

Two stage extrusion of plasticized pectin/poly(vinyl alcohol) blends

Blends of pectin with starch (high amylose and normal), poly(vinyl alcohol) (PVOH), and glycerol were extruded in a twin screw extruder, pelletized, and then further processed into blown film and extruded sheet using a single screw extruder. The samples were analyzed using tensile measurements, dynamic mechanical analysis, and scanning electron microscopy. PVOH levels of 24% or greater were necessary to successfully make blown film, while extruded sheet could be made at a level of 16% PVOH. Tensile strength and initial modulus of the extruded sheets were slightly higher in the machine direction than in the cross direction, while the reverse was true for elongation to break. For the blown films tensile strength tended to be higher in the transverse direction than in the machine direction, while the reverse was seen for initial modulus. Increased levels of PVOH led to increases in tensile strength and elongation to break, while initial modulus was decreased. Morphology as determined by SEM visually indicated stretching in the transverse direction of the blown films. The second stage extrusion appeared to promote -helix formation in the high amylose starch.     

Response of broiler chicks to dietary full-fat soybeans extruded at different temperatures prior to or after grinding

Whole (WSB) or ground (GSB) full-fat soybeans were extruded at 80, 100, 120 or 140°C and incorporated into broiler diets at 30% inclusion level during a 3-week trial.Extrusion of soybeans at 140 °C improved feed intake compared to those extruded at lower temperatures (P < 0.05). However, there were no significant effects of the extrusion temperature on weight gain, feed:gain ratio or mortality rate (P > 0.05). Also, there were no significant effects of whether the extrusion was undertaken prior to or after grinding of the soybeans on feed intake, weight gain, feed:gain ratio, or mortality rate (P > 0.05). There was no significant interaction between the extrusion temperature and the form extruded on any of the performance parameters (P > 0.05).Although increase in extrusion temperature reduced the trypsin inhibitor activity (TIA) of the soybeans, there were no significant effects of the extrusion temperature or type of soybean extruded on nitrogen, fat, calcium or phosphorus retention or diet ME (P > 0.05).It is concluded that low temperature extrusion of full fat soybeans does not reduce TIA to a satisfactory level (< 10 mg g⁻¹ sample) that will enable high dietary inclusion of such extruded soybeans for broilers.     

G790del mutation in DSC2 alone is insufficient to develop the pathogenesis of ARVC in a mouse model

BackgroundArrhythmogenic right ventricular cardiomyopathy (ARVC) is an inherited heart disease that causes heart failure and/or sudden cardiac death. Several desmosomal genes (DSC2, PKG, PKP2, DSP, and RyR2) are thought to be the causative gene involved in ARVC. Out of them, DSC2 mutations account for 2% of ARVC genetic abnormalities. This study aimed to clarify the effect of G790del mutation in DSC2 on the arrhythmogenic mechanism and cardiac function in a mouse model.ResultNeither the heterozygous +/G790del nor homozygous G790del/G790del mice showed structural and functional defects in the right ventricle (RV) or lethal arrhythmia. The homozygous G790del/G790del 6-month-old mice slightly showed left ventricular (LV) dysfunction. Cell shortening decreased with prolongation of intracellular Ca2+ transient in cardiomyocytes isolated from the homozygous G790del/G790del mice, and spontaneous Ca²⁺ transients were frequently observed in response to isoproterenol.ConclusionsG790del mutation in DSC2 was not relevant to the pathogenesis of ARVC, but showed a slight contractile dysfunction and Ca²⁺ dysregulation in the LV.     

Studies on in vitro Extrusion and Ultrastructure of the Spermatophore in Haemaphysalis longicornis (Acari: Ixodidae)

Copulation in ticks is completed by the insertion of the spermatophore into the female genital aperture by a male. The endospermatophore, a cord-like structure and contents are packed in the ectospermatophore of the completed spermatophore. The endospermatophore extrudes just after insertion of the tip of the spermatophore. Only the endospermatophore enters the female genital tract, and the ectospermatophore remains outside the female body. The extrusion is observed in vitro in Haemaphysalis longicornis at various concentrations of NaCl solution: the process is accelerated in less concentrated solutions. The cord-like structure and the endospermatophore finally receive contents extruded from the ectospermatophore. The tip of the cord-like structure connects to the surface of the endospermatophore, and together form a loop after extrusion. Ultrastructural observations confirmed that the ectospermatophore wall is composed of four layers, and the contents consist of male germ cells and three types of secretions from the male accessory genital glands. As in other ticks the male germ cells are elongated spermatids in spermatophores just after formation and extrusion. Adlerocysts described in other ticks are not found in the spermatophore of H. longicornis.     

Effects of Low Moisture Extrusion on the Structural and Physicochemical Properties of Adlay (Coix lacryma-jobi L.) Starch-Based Polymers

This study investigated the changes in the structural and functional properties of various adlay starch-based polymers that occur during low moisture extrusion. The water absorption index, water solubility index, color, and morphological, pasting, and thermal properties of the adlay flours were investigated. Extrusion significantly decreased their contents (starch > protein > fiber > lipids). The color of the adlay flour darkened after extrusion, primarily due to the Maillard reaction and caramelization. Scanning electron microscopy revealed various cracks, pits, and holes on the surface of the extruded flour without defatting for both the extruded brown adlay and the extruded polished adlay. Extrusion caused molecular degradation of the extruded starch, decreasing its swelling power in hot water. The ΔH values (0.19-0.71 J/g) of the extrudates revealed that the starch granules were not fully gelatinized. These results provide a theoretical basis for further explorations of the interactions of adlay macromolecules by extrusion.     

A novel technique for ligation of the cephalic vein reduces hemorrhaging during a two-in-one insertion of dual cardiac device leads

The cutdown technique for the cephalic vein is a common access route for transvenous cardiac device leads (TVLs), and sometimes one cephalic vein can accomodate two TVLs. We examined a novel ligation technique to balance the hemostasis and lead maneuverability for this two-in-one insertion. A total of 22 patients scheduled for cardiac device implantations with two or more leads were enrolled. The ipsilateral cephalic vein was identified for inserting the TVLs with a cutdown. If two TVLs could be introduced into one cephalic vein, hemostasis was established by ligating the venous wall between the TVLs. We measured the amount of hemorrhaging per minute and the operators assessed the lead maneuverability before and after the ligation. We successfully implanted cardiac devices in 15 patients (68%) with this novel method, whereas only one TVL could be introduced via the cephalic vein in 7 patients. As for the successful patients, hemorrhaging from the gap was significantly reduced (5.6?±?7.3 to 0.41?±?0.36g/min, p?=?0.016) after the novel ligation. The lead maneuverability was well maintained so there was no difficulty placing the leads into the cardiac chambers in all cases. No major complications were observed. In the present study, the novel ligation method provided significant hemostasis as well as a preserved maneuverability. It could be an optional choice for insertion of multiple TVLs.     

Extrusions are phagocytosed and promote Chlamydia survival within macrophages

The precise strategies that intracellular pathogens use to exit host cells have a direct impact on their ability to disseminate within a host, transmit to new hosts, and engage or avoid immune responses. The obligate intracellular bacterium Chlamydia trachomatis exits the host cell by two distinct exit strategies, lysis and extrusion. The defining characteristics of extrusions, and advantages gained by Chlamydia within this unique double‐membrane structure, are not well understood. Here, we define extrusions as being largely devoid of host organelles, comprised mostly of Chlamydia elementary bodies, and containing phosphatidylserine on the outer surface of the extrusion membrane. Extrusions also served as transient, intracellular‐like niches for enhanced Chlamydia survival outside the host cell. In addition to enhanced extracellular survival, we report the key discovery that chlamydial extrusions are phagocytosed by primary bone marrow‐derived macrophages, after which they provide a protective microenvironment for Chlamydia. Extrusion‐derived Chlamydia staved off macrophage‐based killing and culminated in the release of infectious elementary bodies from the macrophage. Based on these findings, we propose a model in which C. trachomatis extrusions serve as “trojan horses” for bacteria, by exploiting macrophages as vehicles for dissemination, immune evasion, and potentially transmission.     

Insertion and hairpin formation of membrane proteins: a Monte Carlo study.

Some particular effects of a lipid membrane on the partitioning and the concomitant folding processes of model proteins have been investigated using Monte Carlo methods. It is observed that orientational order and lateral density fluctuations of the lipid matrix stabilize the orientation of helical proteins and induce a tendency of spontaneous formation of helical hairpins for helices longer than the width of the membrane. The lateral compression of the lipids on a hairpin leads to the extrusion of a loop at the trans side of the membrane. The stability of the hairpin can be increased by the design of appropriate groups of hydrophilic and hydrophobic residues at the extruded loop. It is shown that in the absence of lipids the orientation of proteins is not stable and the formation of hairpins is absent. Some analogies between the formation of helical hairpins in membranes and the formation of hairpins in polymer liquid crystals are discussed. The simulations indicate that the insertion process follows a well-defined pattern of kinetic steps.     

Efficient Encapsulation of Plasmid DNA in Anionic Liposomes by a Freeze/Thaw-Extrusion Procedure

Abstract

In this study we investigated whether intact plasmid DNA can be efficiently encapsulated in anionic liposomes prepared by freeze/thaw and extrusion techniques. There is controversy about this method of DNA encapsulation, especially as to whether DNA remains intact and retains its biological activity during extrusion. A solution containing supercoiled plasmid pCMVβ (7164 base pairs) was added to dry lipid films, and after freezing and thawing, the suspension was extruded through a filter with 0.2 μm pores. About 20% of the DNA became encapsulated, as evidenced by protection from degradation by endonuclease added externally. Plasmid isolated from the liposomes was structurally intact, and had essentially the same transfection activity as untreated DNA. These results show that plasmid DNA can be reliably and efficiently encapsulated in anionic liposomes by freeze/thaw and extrusion.