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A temperature-sensitive NUP116 null mutant forms a nuclear envelope seal over the yeast nuclear pore complex thereby blocking nucleocytoplasmic traffic.

NUP116 encodes a 116-kD yeast nuclear pore complex (NPC) protein that is not essential but its deletion (nup116 delta) slows cell growth at 23 degrees C and is lethal at 37 degrees C (Wente, S. R., M. P. Rout, and G. Blobel. 1992. J. Cell Biol. 119:705-723). Electron microscopic analysis of nup116 delta cells shifted to growth at 37 degrees C revealed striking perturbations of the nuclear envelope: a double membrane seal that was continuous with the inner and outer nuclear membranes had formed over the cytoplasmic face of the NPCs. Electron-dense material was observed accumulating between the cytoplasmic face of these NPCs and the membrane seal, resulting in "herniations" of the nuclear envelope around individual NPCs. In situ hybridization with poly(dT) probes showed the accumulation of polyadenylated RNA in the nuclei of arrested nup116 delta cells, sometimes in the form of punctate patches at the nuclear periphery. This is consistent with the electron microscopically observed accumulation of electron-dense material within the nuclear envelope herniations. We propose that nup116 delta NPCs remain competent for export, but that the formation of the membrane seals over the NPCs blocks nucleocytoplasmic traffic.

Animals↗

Respiratory chemosensitivity during wake and sleep in harbour seal pups (Phoca vitulina richardsii).

In this study, we examined the cardiorespiratory patterns of harbour seal pups under normoxic/normocarbic (air), hypoxic/normocarbic (15%, 12%, and 9% O2 in air), and normoxic/hypercarbic (2%, 4%, and 6% CO2 in air) conditions while awake and sleeping on land. Animals were chronically instrumented to record electroencephalogram (EEG), electromyogram (EMG), and electrocardiogram (EKG) signals, which, along with respiration (whole-body plethysmography) and oxygen consumption (VO2), were recorded from animals breathing each gas mixture for 2-4 h on separate days. Our results show that for animals breathing air, VO2 was not significantly lower during slow-wave sleep (SWS; 7.71 +/- 0.39 mL O2 min(-1) kg(-1); all measurements are mean +/- SEM) than during wakefulness (WAKE; 8.80 +/- 0.25 mL O2 min(-1) kg(-1)) and was unaffected by changes in respiratory drive. Although there was no significant fall in VO2 associated with a decrease in arousal state, breathing frequency (f(R)) did decrease (from 18.80 +/- 1.50 breaths min(-1) in WAKE to 10.40 +/- 0.49 breaths min(-1) in SWS), while the incidence of long apneas (>20 s) increased (12.76 +/- 4.06 apneas h(-1) in WAKE and 31.95 +/- 2.37 apneas h(-1) in SWS). Breathing was rarely seen during rapid eye movement (REM) sleep. Tachypnea was present at all levels of increased respiratory drive; however, hypoxia induced a dramatic bradycardia regardless of arousal state, while hypercarbia produced a tachycardia in SWS only. The hypoxic and hypercarbic chemosensitivities of harbour seal pups were similar to those of terrestrial mammals; however, unlike terrestrial mammals, where hypoxic and hypercarbic sensitivities are often reduced during SWS, the sensitivity of harbour seal pups to hypoxia and hypercarbia remained unchanged during the decrease in arousal state from WAKE to SWS.

Analysis of Variance↗

Development of body oxygen stores in harbor seals: effects of age, mass, and body composition.

Harbor seal pups are highly precocial and can swim and dive at birth. Such behavioral maturity suggests that they may be born with mature body oxygen stores or that stores develop quickly during the nursing period. To test this hypothesis, we compared the blood and muscle oxygen stores of harbor seal pups, yearlings, and adults. We found that pups had smaller oxygen stores than adults (neonates 57%, weaned pups 75%, and yearlings 90% those of adults), largely because neonatal myoglobin concentrations were low (1.6+/-0.2 g% vs. 3.8+/-0.3 g% for adults) and changed little during the nursing period. In contrast, blood oxygen stores were relatively mature, with nursing pups having hematocrit (55%+/-0.2%), hemoglobin (21.7+/-0.4 g%), and blood volume (12.3+/-0.5 mL/kg) only slightly lower than the corresponding values for adults (57%+/-0.2%, 23.8+/-0.3 g %, and 15.0+/-0.5 mL/kg). Because neonatal pups had relatively high metabolic rates (11.0 mL O2/kg min), their calculated aerobic dive limit was less than 50% that of adults. These results suggest that harbor seals' early aquatic activity is primarily supported by rapid development of blood, with immature muscle oxygen stores and elevated use rates limiting aerobic diving ability.

Age Factors↗

Evaluation of pressure changes in a new design tracheal tube cuff, the Portex Soft Seal, during nitrous oxide anaesthesia.

We have measured pressure changes in a newly designed tracheal tube cuff, the Portex Soft Seal, during nitrous oxide anaesthesia compared with a Mallinckrodt Lo-Contour tube and a Portex Profile tube. The pressure increases in both control groups were significantly greater than those with the new design (P < 0.0001 in each case). The mean increase in pressure in the Mallinckrodt Lo-Contour tube cuff was 9.9 (SD 3.4) mm Hg compared with 10.3 (1.8) mm Hg in the Portex Profile tube cuff and 2.1 (1.5) mm Hg in the Portex Soft Seal tube cuff. We conclude that the Portex Soft Seal cuff prevented a significant increase in intracuff pressure during nitrous oxide anaesthesia.

Adolescent↗

Interim storage of spent and disused sealed sources: optimisation of external dose distribution in waste grids using the MCNPX code.

Radioactive sealed sources are in use worldwide in different fields of application. When no further use is foreseen for these sources, they become spent or disused sealed sources and are subject to a specific waste management scheme. Portugal does have a Radioactive Waste Interim Storage Facility where spent or disused sealed sources are conditioned in a cement matrix inside concrete drums and following the geometrical disposition of a grid. The gamma dose values around each grid depend on the drum's enclosed activity and radionuclides considered, as well as on the drums distribution in the various layers of the grid. This work proposes a method based on the Monte Carlo simulation using the MCNPX code to estimate the best drum arrangement through the optimisation of dose distribution in a grid. Measured dose rate values at 1 m distance from the surface of the chosen optimised grid were used to validate the corresponding computational grid model.

Computer Simulation↗

New ultrathin-walled endotracheal tube with a novel laryngeal seal design. Long-term evaluation in sheep.

BACKGROUND: A new endotracheal tube (ETT) was fabricated and tested in sheep. It had no tracheal cuff; airway seal was achieved at the level of the glottis through a no-pressure seal made of "gills"; the laryngeal portion was oval-shaped; and the wall thickness was reduced to 0.2 mm. METHODS: Sheep were tracheally intubated either with a standard tube or with the new tube, and their lungs were mechanically ventilated for 1 or 3 days. Air leak was recorded at different peak inspiratory pressures (PIPs). Liquid seepage into the trachea was assessed using an indicator dye. Tracheolaryngeal lesions were scored grossly and histologically. RESULTS: There was no air leak up to 40 cmH2O of PIP, in either group, in short- and long-term studies. Methylene blue leaked across the cuff in two sheep with standard ETTs. No dye leaked across the gills with the new ETTs. In the new ETT group, the trachea appeared better preserved, grossly and histologically, than in the standard ETT group at both 1 and 3 days (P < 0.05). At day 1, the larynx and vocal cords appeared grossly less injured in the new ETT group (P < 0.05), whereas there was no difference at day 3. Histology did not show significant difference on vocal cords, epiglottis, and larynx between the two groups at any time. CONCLUSIONS: The novel, no-pressure seal design of the new ETT is highly effective in preventing air leak and aspiration. It causes no significant tracheal injury.

Animals↗

Tracheal mucus velocity remains normal in healthy sheep intubated with a new endotracheal tube with a novel laryngeal seal.

BACKGROUND: Tracheal mucus velocity (TMV), an index of mucociliary clearance, is reduced markedly in patients intubated with standard endotracheal tubes (ETTs) with high-compliance low-pressure (hi-lo) cuffs. The authors developed a new ultra-thin walled ETT in which the inflatable cuff is replaced with a no-pressure seal, positioned at the level of the larynx. The seal consists of 12 to 20 toroidal layers of thin polyurethane film ("gills") at the level of the vocal cords and prevents both air leak and fluid aspiration. The authors hypothesized that ETTs with the new laryngeal seal may impair TMV less than ETTs with inflated hi-lo cuffs do. METHODS: The TMV was measured in seven healthy female sheep by radiographically tracking the motion of small discs of tantalum inserted into the trachea through a bronchoscope. The TMV was measured in spontaneously breathing sheep before intubation (baseline) and after intubation with either a hi-lo ETT (control group) or after intubation with a new ETT with gills (study group). Four to six weeks later, the studies were repeated, but the sheep that were previously in the control group served as the study group, and those in the study group served as controls. RESULTS: Baseline TMV did not differ in the two groups. In the control group, TMV decreased significantly (by 67%) from baseline. In the study group, TMV did not differ significantly from baseline and remained steady during 3 h of intubation. CONCLUSIONS: The TMV does not change in sheep intubated with new ETTs with gills. The new ETT's may help promote a normal mucociliary clearance in patients who require ventilation.

Animals↗

The performance of commercially available sealed Dacron vascular grafts in intraarterial thrombolytic therapy.

The purpose of this study was to assess whether two commercially available, sealed arterial prostheses would tolerate catheter directed, intraarterial thrombolytic therapy. Sealed vascular grafts were implanted in 12 mongrel dogs for 3, 4, and 6 week intervals as iliac artery interposition grafts. This allowed direct comparison of two types of sealed grafts in the same animal. The indications for graft failure consisted of clinically evident bleeding, transluminal extravasation, or anastomotic leakage of contrast medium documented by angiography. Thrombolytic therapy (urokinase) was delivered immediately proximal to the grafts in the distal aorta, thereby exposing the intraluminal surfaces to urokinase delivered directly from the catheter tip, as well as to the accumulated systemic levels of urokinase and urokinase activated plasmin. There was no evidence of postoperative bleeding or extravasation of contrast medium during any follow-up arteriogram in any animal. There was no discernible extravasation of contrast medium noted at the anastomotic sites. At the time of explantation, no graft demonstrated any evidence of subclinical extravasation, such as separation of the external capsule from the abluminal surface of the graft. It is concluded that it appears safe to use thrombolytic therapy in these grafts as early as 3 weeks postimplantation.

Animals↗

Evaluation of a new bioadhesive copolymer (ADAL) to seal corneal incisions.

OBJECTIVE: To evaluate the efficacy and clinical tolerance of a new acrylic copolymer tissue adhesive, ADAL, in corneal surgery. METHODS: Corneal incisions were performed on 24 New Zealand albino rabbits. The incision was either sealed with ADAL bioadhesive, hydrated with balanced salt solution, or closed with 10-0 nylon surgical sutures. The incisions underwent weekly tensile strength testing and clinical, histopathologic, and confocal microscopy evaluations. RESULTS: Clinical evaluation revealed good sealing and no incision leakage in any group. There was no anterior chamber reaction in any case. Compared with the hydrated group, there was somewhat more incisional vascularization and localized corneal opacity in the ADAL group, but there were no cases of severe corneal opacity or neovascularization. On histologic analysis, a slightly larger inflammatory reaction was noted in the ADAL group compared with the hydrated group. Tensile strength during the first week was statistically significantly greater in the ADAL group (2.93 Newtons) than in the control groups (1.40 Newtons, hydrated group; 1.43 Newtons, sutured group) (P < 0.001). Confocal microscopy study revealed a unique dendritic keratocyte shape and structure resembling shiny crystalline particles after disappearance of the adhesive from the cornea. CONCLUSIONS: Corneal surgical incisions sealed with ADAL adhesive have superior tensile strength in the first postoperative week compared with incisions closed with sutures or hydrated with salt solution. Thereafter, findings in both groups became similar. The use of ADAL adhesive was not associated with any deleterious effects during the corneal incision-healing process compared with hydration or sutures.

Acrylates↗

The effect of immediate vs. delayed post space preparation on the apical seal of a root canal filling: a study in an increased-sensitivity pressure-driven system.

A 5 mm remaining length of root canal filling, after post space preparation, is commonly assumed to maintain sealing ability similar to that of the intact filling. Post spaces were prepared either immediately using hot pluggers, or later, using drills. The sealing ability of the fillings, 5 mm remaining length, were compared with each other and with an intact root canal filling control, using radioactive tracer in a pressure-driven system. When no pressure was applied, no differences could be detected between either of the groups and the control. When a pressure of 120 mm Hg was applied to the same teeth, the control group clearly maintained a better seal than each of the experimental groups, which did not significantly differ from each other. These results suggest that (a) the pressure-driven system was more sensitive than the passive leakage assay that failed to detect differences even at 14 days; (b) a remaining root canal filling of 5 mm was inferior to the intact root canal filling; and (c) the immediate post space preparation with hot pluggers did not differ from a delayed preparation with drills.

Analysis of Variance↗

Effect of three different sealers on the sealing ability of both thermafil obturators and cold laterally compacted Gutta-Percha.

The purpose of this study was to assess the seal obtained in straight and curved root canals filled with either laterally compacted gutta-percha or Thermafil obturators. Each technique was used in combination with three different sealers (RSA RoekoSeal, AH Plus, AH 26). Thermafil obturators were also used without sealer, resulting in a total of 14 test groups of 16 teeth each. Sixty teeth served as positive or negative controls. Included in the study were 142 extracted teeth with straight and 142 with curved root canals. All canals were enlarged up to size 40. After obturation, all roots were placed in India ink for 48 h and rendered transparent to measure the maximum linear dye penetration. Canals filled with Thermafil obturators had significantly more extrusion of filling material than canals filled by lateral compaction (p < 0.01). Thermafil without sealer showed significantly greater dye penetration compared with all other groups both in straight and in curved canals (p < 0.05). As long as a sealer was used, the seal obturated with Thermafil was equivalent in terms of dye penetration to lateral compaction. There were no statistical differences in the mean apical dye penetration among the three sealers. The differences between the dye penetration in straight and in curved canals were insignificant for all groups (p > 0.05). Under the conditions of this study, Thermafil obturators achieved seals comparable to lateral compaction, as long as a sealer was used.

Analysis of Variance↗

Detachable self-sealing membrane system for the endoscopic treatment of incontinence.

PURPOSE: The endoscopic treatment of urinary incontinence is limited by the injectable substances currently available. The ideal injectable material should be able to conserve its volume, be nonmigratory and nonantigenic. Toward this goal we developed a system for the treatment of incontinence in which a catheter with an inflatable and detachable self-sealing silicone membrane fits through a standard cystoscopic needle. We present our experience with this system. MATERIALS AND METHODS: A total of 20 female beagle dogs underwent cystoscopy and in 15 a self-sealing membrane was placed endoscopically in the submucosal region of the proximal urethra at the 3 and 9 o'clock positions. Each membrane was inflated through the delivery catheter with 0.2 cc povidone. Coaptation of the urethra due to the relative bulking effect of the inflated membrane was confirmed endoscopically in each animal. Five control animals received only an injection of saline. Four animals were sacrificed at 1, 3, 6, 12 and 18 months, respectively. At sacrifice gross and histological examinations were performed. RESULTS: At retrieval up to 18 months after implantation the membrane remained inflated and at the same position at which it had been initially placed. There was no evidence of povidone volume loss or extravasation. Membranes were encapsulated by a fibrous capsule. Only a few inflammatory cells surrounded the capsule by month 1 and there were none by month 3. Histological examination of periurethral tissue and distant organs showed no evidence of particle migration. CONCLUSIONS: The detachable self-sealing membrane system is easily implantable, nonmigratory, nonantigenic and able to conserve its volume. These studies indicate that this system may be effective for the endoscopic treatment of incontinence.

Animals↗

Use of the LigaSure vessel sealing device in laparoscopic living-donor nephrectomy.

BACKGROUND: Laparoscopic living-donor nephrectomy has gained acceptance within the transplant community. The technique requires advanced laparoscopic techniques, and great care must be taken to ensure safety of the operation for the donor and transplantability of the donor kidney. Minimizing the risk of bleeding and trauma to the kidney are important features of a successful living-donor nephrectomy. Improved laparoscopic instrumentation has afforded greater safety and efficacy through technical advances. METHODS: The LigaSure device was used in 124 consecutive living-donor nephrectomies beginning in 1999. A transplant database was reviewed for operative statistics including intraoperative blood loss and operating time. RESULTS: The LigaSure device was used to dissect and seal all venous and arterial branches. Estimated blood loss was 90 +/- 53 mL. A suction device was required in only 40 (32%) of the cases. No patient experienced postoperative bleeding. There were two donor complications: one incisional hernia and one ileus. All kidneys functioned immediately upon reperfusion. CONCLUSIONS: The LigaSure device is an extremely effective tool for obtaining hemostasis by sealing both venous and arterial branches of the major renal vessels. This is also effective in sealing lymphatic tissues and thereby facilitating dissection. Avoiding the use of metallic clips simplified final division of the renal artery and vein. As with any laparoscopic instrument, the anatomic geometry of the operative field may limit use based upon port placement.

Adolescent↗

Sealing ability of amalgam used as a retrograde root filling in endodontic surgery.

A dye penetration technique was used to investigate the sealing ability of amalgam retrograde root fillings in vitro. A factorial design was employed to determine the effect of amalgam fillings of 1, 2 and 4 mm in length with and without prior filling of the canals with gutta-percha. The results were compared with a group of teeth filled with laterally condensed gutta-percha alone and another with laterally condensed gutta-percha plus root resection but with no retrograde filling. It was concluded that increasing the length of the amalgam filling did not improve the seal, that prior filling of the canal did not improve the seal, that retrograde root fillings were no worse than laterally condensed gutta-percha alone and that root resection of teeth filled with laterally condensed gutta-percha without retrograde filling was no worse than those filled with laterally condensed gutta-percha without root resection or retrograde root filling.

Dental Amalgam↗

Sealing ability of materials used as retrograde root fillings in endodontic surgery.

The sealing ability of retrograde root fillings of amalgam plus cavity varnish, EBA cement, glass ionomer cement, light-cured composite resin, dentine bonding agent, and light-cured composite resin plus dentine bonding agent were compared with laterally condensed conventional root fillings. A dye penetration technique was used to assess microleakage and it was concluded that none of the materials produced a perfect seal but that glass ionomer cement, light-cured composite resin, light-cured composite resin plus dentine bonding agent and dentine bonding agent alone produced better seals than conventional laterally condensed gutta-percha, amalgam plus cavity varnish and EBA cement.

Composite Resins↗

Apical and coronal seal of roots obturated with a dentine bonding agent and resin.

The efficacy of sealing root canal systems with a dentine bonding agent and resin cement was compared with that of a glass ionomer sealer. Scanning electron microscopic examination of the bonding agent-dentine interface was also performed. The root canals of 50 single rooted teeth with mature apices were prepared chemomechanically and the smear layer removed with 17% REDTA and 5.25% NaOCl. Specimens were divided into two groups of 24 teeth each. One group had the dentine conditioned with a 10:3 citric acid-ferric chloride solution and obturated with the dentine bonding agent and resin, radiopaque C & B Metabond. The other group was obturated with the glass ionomer sealer, Ketac-Endo and a single cone of gutta-percha. After immersion in Indian ink for 90 h, the teeth were cleared and the quality of the apical and coronal seal was assessed using India ink dye penetration. There was a significantly better seal in both the apical and coronal directions when using the dentine bonding agent and resin obturation material. Scanning electron microscopic examination of the demineralized dentine and the C & B Metabond interface revealed the presence of the characteristic hybrid layer along with microtags of resin penetrating deep into the dentine tubules.

Boron Compounds↗

Fusion of myogenic cells to the newly sealed region of damaged myofibres in skeletal muscle regeneration.

In regenerating skeletal muscle, sarcoplasmic extensions containing variable numbers of nuclei, widely referred to as 'buds' or 'stumps', are formed at the ends of damaged myofibres. In this paper we investigated whether the nuclei seen in the buds results from fusion of myogenic cells or from migration of myonuclei to the sealed ends of damaged myofibres in murine muscle regenerating after crush injury. The fusion of mononuclear and multinucleate myogenic cells to the buds was demonstrated by transmission electron microscopy. In order to elucidate the frequency and kinetics of cytoplasmic continuity between myotubes and sealed myofibres, we labelled the damaged myofibres with carbocyanine dye DiI (which inserts into the lipid bilayer and travels down continuous membranes) and the samples were then examined by confocal scanning microscopy. This technique showed that there was little fusion between myotubes and myofibres during the first 6 days after crush injury, but significant fusion had occurred by the tenth day especially at the newly sealed region of the damaged myofibre. A scheme for the repair of damaged skeletal muscle is presented.

Animals↗

Fit and seal characteristics of a new paediatric tracheal tube with high volume-low pressure polyurethane cuff.

BACKGROUND: To evaluate a new paediatric tracheal tube (Microcuff, Weinheim, Germany) with an ultrathin high volume-low pressure polyurethane cuff. METHODS: With approval of the Hospital Ethics Committee tracheas of children undergoing general anaesthesia were intubated using a Microcuff tube. Tube sizes were selected according to: internal diameter (mm) = age/4 + 3.5 in children aged > or = 2 years. In newborns (> or = 3 kg) < or = 1 year, ID 3.0-mm tubes, and in children from 1 to 2 years, internal diameter 3.5-mm tubes were used. Tubes were classified too large if no air leakage was obtained at an airway pressure of 20 cm H2O with the cuff not inflated. Sealing pressure was assessed by auscultation. Post-extubation croup requiring therapy was noted. RESULTS: Five-hundred children were studied. In eight children the tubes were too large. Sealing pressure was 9.7 +/- 2.5 cm H2O (4-20). In two patients postextubation croup required singular short-term therapy. CONCLUSIONS: Microcuff paediatric tracheal tubes provided tracheal sealing with cuff pressures considerably lower than usually accepted. The rate of tube exchange was very low (1.6%), as was the rate of airway morbidity (croup requiring therapy; 0.4%).

Adolescent↗