Permeabilization of E-coli K12 inner and outer membranes by bothropstoxin-I, A LYS49 phospholipase A2 from Bothrops jararacussu

Although lacking catalytic activity, the Lys49-PLA(2)s damage artificial membranes by a Ca2+-independent mechanism, and demonstrate a potent bactericidal effect. The relationship between the membrane-damaging activity and bactericidal effect of bothropstoxin-I (BthTx-1), a Lys49-PLA(2) from the venom of Bothrops jararacussu, was evaluated for the wildtype protein and a series of site-directed mutants in the active site and C-terminal regions of the protein. The membrane permeabilization effect against the inner and outer membranes of Escherichia coli K12 was evaluated by fluorescence changes of Sytox Green and N-phenyl-N-naphthylamine, respectively. With the exception of H48Q, all mutants reduced the bactericidal activity, which correlated with a reduction of the permeabilization effect both against the inner bacterial membrane. No significant differences in the permeabilization of the bacterial outer membrane were observed between the native, wild-type recombinant and mutant proteins. These results suggest different permeabilization mechanisms against the inner and outer bacterial membranes. Furthermore, the structural determinants of bacterial inner membrane damage identified in this study correlate with those previously observed for artificial membrane permeabilization, suggesting that a common mechanism of membrane damage underlies the two effects. (C) 2007 Elsevier Ltd. All rights reserved.

QUADRANT-SEARCHING: A NEW TECHNIQUE TO REDUCE THE SEARCH SPACE OF THE INVERSE PROBLEM OF EIT USING BEM TO THE DIRECT PROBLEM

The image reconstruction using the EIT (Electrical Impedance Tomography) technique is a nonlinear and ill-posed inverse problem which demands a powerful direct or iterative method. A typical approach for solving the problem is to minimize an error functional using an iterative method. In this case, an initial solution close enough to the global minimum is mandatory to ensure the convergence to the correct minimum in an appropriate time interval. The aim of this paper is to present a new, simple and low cost technique (quadrant-searching) to reduce the search space and consequently to obtain an initial solution of the inverse problem of EIT. This technique calculates the error functional for four different contrast distributions placing a large prospective inclusion in the four quadrants of the domain. Comparing the four values of the error functional it is possible to get conclusions about the internal electric contrast. For this purpose, initially we performed tests to assess the accuracy of the BEM (Boundary Element Method) when applied to the direct problem of the EIT and to verify the behavior of error functional surface in the search space. Finally, numerical tests have been performed to verify the new technique.

The outer wall of small airways is a major site of remodeling in fatal asthma

Background: Structural and inflammatory changes in asthma involve both the large and small airways, with involvement of the distal lung being related to disease severity. We have previously shown that changes in the extracellular matrix (ECM) composition of the distal lung are associated with loss of alveolar attachments in patients with fatal asthma. However, major ECM elements, such as collagen I and fibronectin and their regulators, have not been addressed at the distal level. Objective: We sought to evaluate ECM remodeling in the distal lungs of asthmatic patients. Methods: Using immunohistochemistry and image analysis, we determined the content of collagen I and III, fibronectin, and matrix metalloproteinases; (MMPs) 1, 2, and 9 and tissue inhibitors of metalloproteinase (MMPs) 1 and 2 in the large and small airways and lung parenchyma of 24 patients with fatal asthma and compared the results with those of 11 nonasthmatic control subjects. Protein content was defined as the area of positive staining divided by basement membrane or septum length. Results: We observed increased collagen I and decreased collagen III content in the small airways of asthmatic patients compared with that seen in control subjects. Greater fibronectin and MMP-1, MMP-2, and MMP-9 content was observed at the outer area of the small airways in asthmatic patients. NIMP content was also increased in the peribronchiolar parenchyma in asthmatic patients. In contrast, TIMP expression was only increased in the large airways of asthmatic patients compared with that seen in control subjects. Conclusions: The outer area of the small airways is a major site of ECM remodeling in fatal asthma, potentially contributing to functional changes and the loss of airway-parenchyma interdependence observed in patients with fatal asthma. (J Allergy Clin Immunol 2009;123:1090-7.)

Distribution of fentanyl in the placental intervillous space and in the different maternal and fetal compartments in term pregnant women

Fentanyl is used in obstetrical practice to promote analgesia and anesthesia during labor and in cesarean delivery, with rapid and short-term effects. To determine fentanyl concentrations in maternal plasma, in the placental intervillous space, and in the umbilical artery and vein in term pregnant women. Ten healthy pregnant women underwent epidural anesthesia with fentanyl plus bupivacaine and lidocaine, and fentanyl concentrations were determined in the various maternal and fetal compartments, including the placental intervillous space, which has not been previously studied in the literature. The ratios of fentanyl concentrations in the various maternal and fetal compartments revealed an 86% rate of placental fentanyl transfer. The highest fentanyl concentrations were detected in the placental intervillous space, being 2.19 times higher than in maternal plasma, 2.8 times higher than in the umbilical vein and 3.6 times higher than in the umbilical artery, with no significant differences between the umbilical vein and artery, demonstrating that there was no drug uptake by fetal tissues nor metabolism of the drug by the fetus despite the high rates of placental transfer. The present study demonstrated that the placental intervillous space acted as a site of fentanyl deposit, a fact that may be explained by two hypotheses: (1) the blood collected from the placental intervillous space is arterial and, according to some investigators, the arterial plasma concentrations of the drugs administered to patients undergoing epidural anesthesia are higher than the venous concentrations, and (2) a possible role of P-glycoprotein (P-gp).

Distribution of Bupivacaine Enantiomers and Lidocaine and Its Metabolite in the Placental Intervillous Space and in the Different Maternal and Fetal Compartments in Term Pregnant Women

The aim of this study is to determine the concentrations of lidocaine and its metabolite, monoethylglycine xylidide (MEGX), and of the enantiomers of bupivacaine in maternal and fetal compartments. Ten healthy pregnant women were submitted to epidural anesthesia. Drug concentrations were determined in the maternal vein, fetal umbilical artery and vein, and the placental intervillous space. The highest concentrations of the bupivacaine enantiomers lidocaine and of lidocaine and of its MEGX metabolite were detected in maternal plasma and in the placental intervillous space. The placental transfer was 33% for the (+)-(R)-bupivacaine enantiomer and 31% for the (-)-(S)-bupivacaine enantiomer. For lidocaine and its MEGX metabolite, respective placental transfers were 60% and 43%. Lidocaine concentration in the fetal umbilical vein was 1.46 times higher than in the fetal umbilical artery. The highest concentrations of lidocaine and its metabolite and of the enantiomers of bupivacaine were detected in the placental intervillous space. The higher lidocaine concentrations in the fetal umbilical vein than in the fetal umbilical artery suggest that there was tissue uptake of the drug or drug metabolization by the fetus.

Does rapid maxillary expansion increase nasopharyngeal space and improve nasal airway resistance?

Objective: To evaluate the effect of rapid maxillary expansion (RME) on the dimension of the nasopharyngeal space and its relation to nasal airway resistance. Methods: Twenty-five school-age children (from 7 to 10 year-old) with mouth and/or mixed breathing, with mixed dentition and uni- or bilateral posterior crossbite involving the deciduous canines and the first permanent molars, were evaluated. RME was placed and remained during 90 days. Rhinomanometry and orthodontic documentation were performed at four different times, i.e., before (T(1)), immediately after (T(2)), 90 days (T(3)) and 30 months (T(4)) after RME. Results: Differences in nasopharyngeal area and in nasal airway resistance were observed only 30 months after RME, and could be explained by facial growth, and not because of the orthodontic procedure. Conclusion: RME does not influence on nasopharyngeal area or nasal airway resistance in long-term evaluation. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

Spontaneous reversibility of damage to outer hair cells after sodium salicylate induced ototoxicity

Background: High sodium salicylate doses can cause reversible hearing loss and tinnitus, possibly due to reduced outer hair cell electromotility. Sodium salicylate is known to alter outer hair cell structure and function. This study determined the reversibility and cochlear recovery time after administration of an ototoxic sodium salicylate dose to guinea pigs with normal cochlear function. Study design: Prospective experimental investigation. Methods: All animals received a single 500 mg sodium salicylate dose, but with different durations of action. Function was evaluated before drug administration and immediately before sacrifice. Cochleae were processed and viewed using scanning electron microscopy. Results: Changes in outer hair cell function were observed to be present 2 hours after drug administration, with recovery of normal anatomy beginning after 24 hours. Subsequently, derangement and distortion of cilia reduced, with effects predominantly in row three. At 168 hours, cilia were near-normal but with mild distortions which interfered with normal cochlear physiology. Conclusions: Ciliary changes persisted for up to 168 hours after ototoxic sodium salicylate administration.

Percutaneous Transatrial Access to the Pericardial Space for Epicardial Mapping and Ablation

Background-Puncture of the atrial appendage may provide access to the pericardial space. The aim of this study was to evaluate the feasibility of epicardial mapping and ablation through an endocardial transatrial access in a swine model. Methods and Results-An 8-F Mullins sheath was used to perforate the right (n=16) or left (n=1) atrial appendage in 17 pigs (median weight, 27.5 kg; first and third quartiles [Q1, Q3], 25.2, 30.0 kg). A 7-F ablation catheter was introduced into the pericardial space to perform epicardial mapping and deliver radiofrequency pulses on the atria. The pericardial space was entered in all 17 animals. In 15 (88%) animals, there was no hemodynamic instability (mean blood pressure monitoring, initial median, 80 mm Hg; Q1, Q3, 70, 86 mm Hg; final median, 88 mm Hg; Q1, Q3, 80, 96 mm Hg; P=0.426). In these 15, a mild hemorrhagic pericardial effusion was identified and aspirated (median, 20 mL; Q1, Q3, 15, 30 mL) during the procedure, and postmortem gross analysis revealed that the atrial perforation was closed in these animals. In 2 (12%) of the 17 animals, there was major pericardial bleeding with hemodynamic collapse. On gross examination, it was found that pericardial space was accessed through right ventricular perforation in 1 animal and the tricuspid annulus in the other. After the initial study, we used an occlusion device in 3 other animals to attempt to seal the puncture (2 at the right atrial appendage and 1 at the right ventricle). These 3 animals had no significant pericardial bleeding. Conclusions-Transatrial endovascular right atrial appendage puncture may provide a potential alternative route for pericardial access. Further studies are needed to evaluate its safety with longer and more-complex procedures before being applied in clinical settings. (Circ Arrhythm Electrophysiol. 2011;4:331-336.)

Resin cement thickness in oval-shaped canals: Oval vs. circular fiber posts in combination with different tips/drills for post space preparation

Purpose: To evaluate the cement thickness around oval and circular posts luted in oval post spaces prepared with different drills/tips. Methods: Extracted premolars were endodontically treated and obturated, then randomly divided into three groups (n = 5) according to the tips/drills used for post-space preparation and to the type of fiber post luted: medium grit oval tip + oval posts, fine grit oval tip + oval posts, Mtwo Post File drill + circular posts. The specimens were sectioned in horizontal slices; one slice per canal third was chosen for each post-space, resulting in three slices for each specimen. The distances between the canal wall and the post perimeter were measured on SEM images of each slice. Results: The fine grit tip + oval post group obtained statistically significant lower cement thicknesses than the other groups (P < 0.05), in particular in the apical third. The MtwoPF + circular post group showed the highest cement thickness, comparable to that of the medium tip + oval post group. A good post fitting in oval-shaped canals can be obtained using a fine grit oval tip combined with oval posts. (Am J Dent 2009;22:290-294).

Post space debridement in oval-shaped canals: The use of a new ultrasonic tip with oval section

This study evaluates the effect on post space debridement in oval-shaped canals of an experimental ultrasonic tip with oval section (Satelec) compared with a circular ultrasonic tip (KaVo). Thirty teeth with an oval-shaped canal were endodontically treated and obturated and then randomly divided into 3 groups (n = 10) according to the procedure used for post space debridement: Satelec tip, Largo #2 drill + KaVo file, and Largo #2 drill + water. Debris and dentin tubules were evaluated by assigning scores to scanning electron microscope post spaces images; lower scores corresponded to fewer debris and higher number of open tubules. The Satelec group showed significantly lower debris and open tubules scores than KaVo group (p < .05) and control group (p < .05), which differed significantly between each other (p < .05). Also the debris and open tubules scores in different post space regions differed significantly among the experimental groups (p < .001). The oval ultrasonic tip resulted in a better post space debridement than a circular ultrasonic tip in oval-shaped canals.

Post space cleaning using a new nickel titanium endodontic drill combined with different cleaning regimens

This study compared the effect of two drills and five cleaning regimens on post space debridement. One hundred extracted premolars were instrumented and obturated with warm vertical compaction of gutta percha. The teeth were divided into two groups according to the drill used to remove gutta percha/sealer and for post space preparation: a Largo drill (Largo; Dentsply, St Quentin en Yvelines, France) or a MTwo-PF drill (Sweden&Martina, Due Carrare, Padova, Italy). The following cleaning regimens were used: EDTA, ultrasonics, ultrasonics + EDTA, phosphoric acid, and distilled water. Scanning electron microscopic images of the post spaces were taken, and the presence of debris and of open dentin tubules were evaluated. The ultrasonics + EDTA, phosphoric acid, and EDTA groups were comparable in open tubules scores for both drills and in debris scores after the use of MTwo-PF (p > 0.05). The ultrasonics and control groups performed significantly worse (p < 0.05). The MTwo-PF drill resulted as effective as the Largo drill in obtaining a good post space cleaning, especially when followed by ultrasonics + EDTA irrigant regimen.

Histological Outcomes on the Development of New Space-making Devices for Maxillary Sinus Floor Augmentation

Background: Previous studies have pointed out that the mere elevation of the maxillary sinus membrane promotes bone formation without the use of augmentation materials. Purpose: This experimental study aimed at evaluating if the two-stage procedure for sinus floor augmentation could benefit from the use of a space-making device in order to increase the bone volume to enable later implant installation with good primary stability. Materials and Methods: Six male tufted capuchin primates (Cebus apella) were subjected to extraction of the three premolars and the first molar on both sides of the maxilla to create an edentulous area. The sinuses were opened using the lateral bone-wall window technique, and the membrane was elevated. One resorbable space-making device was inserted in each maxillary sinus, and the bone window was returned in place. The animals were euthanatized after 6 months, and biopsy blocks containing the whole maxillary sinus and surrounding soft tissues were prepared for ground sections. Results: The histological examination of the specimens showed bone formation in contact with both the schneiderian membrane and the device in most cases even when the device was displaced. The process of bone formation indicates that this technique is potentially useful for two-stage sinus floor augmentation. The lack of stabilization of the device within the sinus demands further improvement of space-makers for predictable bone augmentation. Conclusions: It is concluded that (1) the device used in this study did not trigger any important inflammatory reaction; (2) when the sinus membrane was elevated, bone formation was a constant finding; and (3) an ideal space-making device should be stable and elevate the membrane to ensure a maintained connection between the membrane and the secluded space.

Unilateral Correction of Space Loss in the Mixed Dentition

Distalization of maxillary molars is indicated for correction of Class II dental malocclusion and for space gain in cases of space deficiency. The ideal treatment with an intraoral fixed appliance for molar distalization should fulfill the following requirements: patient compliance; acceptable esthetics; comfort; minimum anterior anchor loss (as evidenced by inclination of incisors); bodily movement of the molars to avoid undesirable effects and unstable outcomes; and minimum time required during sessions for placement and activations. The purpose of this paper was to present an alternative treatment for space recovery in the area of the maxillary right second premolar when there has been significant mesial movement of the permanent maxillary right first molar. We used a modified appliance that allows unilateral molar distalization in cases of unilateral tooth/arch size discrepancy using the opposite side as anchor, thus reducing the mesialization of the anterior teeth. (Pediatr Dent 2008;30:334-41) Received August 17, 2006 / Last Revision October 17, 2007 / Revision Accepted October 17, 2007

Contribution to the ground-based follow-up of the Gaia space mission

The Gaia Space Mission [Mignard, F., 2005. The three-dimensional universe with Gaia. ESA/SP-576; Perryman, M., 2005. The three-dimensional universe with Gaia. ESA/SP-576] will observe several transient events as supernovae, microlensing, gamma ray bursts and new Solar System objects. The satellite, due to its scanning law, will detect these events but will not be able to monitor them. So, to take these events into consideration and to perform further studies it is necessary to follow them with Earth-based observations. These observations could be efficiently done by a ground-based network of well-equipped telescopes scattered in both hemispheres. Here we focus our attention at the new Solar System objects to be discovered and observed by the Gaia satellite [Mignard, F., 2002. Observations of Solar System objects by Gaia I. Detection of NEOS. Astron. Astrophys. 393, 727] mainly asteroids, NEOs and comets. A dedicated ground-based network of telescopes as proposed by Thuillot [2005. The three-dimensional universe with Gaia. ESA/SP-576] will allow to monitor those events, to avoid losing them and to perform a quick characterization of some physical properties which will be important for the identification of these objects in further measurements by Gaia. We present in this paper, the beginning of the organization of a Latin-American ground-based network of telescopes and observers joining several institutions in Argentina, Bolivia, Brazil and other Latin-American countries aiming to contribute to the follow-up of Gaia science alerts for Solar System objects. (C) 2008 Elsevier Ltd. All rights reserved.

Relativistic dispersion effects on SS433 jets

We here investigate the dispersion properties of radiation in the SS433 relativistic jets. We assume that the jet is composed of cold electron-proton plasma immersed in a predominantly parallel magnetic field to the jet axis. We find that for the mildly relativistic source SS433 (for which similar or equal to 79 degrees), the bulk velocity is too small (v similar or equal to 0.26c) to produce significant changes in the dispersion properties of the medium. Nonetheless, in the rarefied outer regions of the jets, where radio emission dominates, even a weak magnetic field has some influence on the dispersion properties and there appear two different electromagnetic branches that are slightly sensitive to the bulk relativistic motion. In the inner, X-ray region, the magnetic field is much stronger, but in this region the high electron density preserves the isotropic character of the local plasma and no branch separation occurs. In the region of the jet where the IR and optical emission dominates, the cold plasma may be also considered isotropic, i.e., neither the magnetic field nor the bulk velocity is able to affect the propagation of the radiation. Finally, we find that the Doppler line displacement in SS433 is affected by plasma dispersion only in a narrow frequency range in the far IR. As a consequence, although the shift (z) modulation due to precession of the SS433 jets is well described by previous work, it has to be corrected by plasma dispersion effects in the far-IR range.