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Vivo Experiments (vivo + experiment)
Kinds of Vivo Experiments Selected AbstractsAnimal Research: Reporting In Vivo Experiments: The ARRIVE guidelinesEXPERIMENTAL PHYSIOLOGY, Issue 8 2010Article first published online: 9 JUL 2010 No abstract is available for this article. [source] The ARRIVE guidelines, a welcome improvement to standards for reporting animal researchTHE JOURNAL OF GENE MEDICINE, Issue 7 2010Olivier Danos Abstract Here we introduce the ARRIVE (Animal Research: Reporting In Vivo Experiments) guidelines, produced by the National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), which are published in this issue of the journal with our endorsement, and will be incorporated into our Instructions to Authors. Copyright © 2010 John Wiley & Sons, Ltd. [source] Animal Research: Reporting In Vivo Experiments: The ARRIVE guidelinesTHE JOURNAL OF PHYSIOLOGY, Issue 14 2010Article first published online: 16 JUL 2010 No abstract is available for this article. [source] Guidelines for reporting experiments involving animals: the ARRIVE guidelinesBRITISH JOURNAL OF PHARMACOLOGY, Issue 7 2010JC McGrath British Journal of Pharmacology (BJP) is pleased to publish a new set of guidelines for reporting research involving animals, simultaneously with several other journals; the ,ARRIVE' guidelines (Animals in Research: Reporting In Vivo Experiments). This editorial summarizes the background to the guidelines, gives our view of their significance, considers aspects of specific relevance to pharmacology, re-states BJP's guidelines for authors on animal experiments and indicates our commitment to carrying on discussion of this important topic. We also invite feedback via the British Pharmacological Society website. [source] Reconstruction of MR images from data acquired on an arbitrary k -space trajectory using the same-image weightMAGNETIC RESONANCE IN MEDICINE, Issue 2 2002Yongxian Qian Abstract A sampling density compensation function denoted "same-image (SI) weight" is proposed to reconstruct MR images from the data acquired on an arbitrary k -space trajectory. An equation for the SI weight is established on the SI criterion and an iterative scheme is developed to find the weight. The SI weight is then used to reconstruct images from the data calculated on a random trajectory in a numerical phantom case and from the data acquired on interleaved spirals in an in vivo experiment, respectively. In addition, Pipe and Menon's weight (MRM 1999;41:179,186) is also used in the reconstructions to make a comparison. The images obtained with the SI weight were found to be slightly more accurate than those obtained with Pipe's weight. Magn Reson Med 48:306,311, 2002. © 2002 Wiley-Liss, Inc. [source] Green tea extract weakens the antibacterial effect of amoxicillin in methicillin-resistant Staphylococcus aureus infected micePHYTOTHERAPY RESEARCH, Issue 1 2010Qing Peng Abstract Tea (Camellia sinensis) has been known for its modulation of resistance of methicillin-resistant Staphylococcus aureus (MRSA) to ,-lactam antibiotics in vitro. This study aimed to confirm the in vitro effect of green tea extracts with ,-lactams and to determine whether green tea extracts can reduce the minimum inhibitory concentrations (MICs) of amoxicillin in MRSA-infected mice. The catechins in the test tea that account for the reduced resistance to ,-lactams were quantitatively determined by high-performance liquid chromatography. The MICs of the ampicillin, cefazolin, amoxicillin, oxacillin, tea extract alone and tea extract in combination with ,-lactams were determined. Proportions of tea extracts and amoxicillin-tea extract combinations were administered to groups of mice enterally. The in vitro experiment showed that the MICs of four ,-lactams were greatly decreased in the presence of 0.25% tea extract. However, in an in vivo experiment, amoxicillin in combination with 5% tea extract conferred a higher ED50 than that of antibiotic alone. Green tea extract, alone or in combination with amoxicillin, does not have protective benefits in MRSA-infected mice. This study concluded that tea extract weakened the antibacterial effect of amoxicillin in MRSA infected mice. Tea drinking is not recommended in combination with amoxicillin treatment. Copyright © 2009 John Wiley & Sons, Ltd. [source] Three-dimensional ultrasound image-guided robotic system for accurate microwave coagulation of malignant liver tumoursTHE INTERNATIONAL JOURNAL OF MEDICAL ROBOTICS AND COMPUTER ASSISTED SURGERY, Issue 3 2010Jing Xu Abstract Background The further application of conventional ultrasound (US) image-guided microwave (MW) ablation of liver cancer is often limited by two-dimensional (2D) imaging, inaccurate needle placement and the resulting skill requirement. The three-dimensional (3D) image-guided robotic-assisted system provides an appealing alternative option, enabling the physician to perform consistent, accurate therapy with improved treatment effectiveness. Methods Our robotic system is constructed by integrating an imaging module, a needle-driven robot, a MW thermal field simulation module, and surgical navigation software in a practical and user-friendly manner. The robot executes precise needle placement based on the 3D model reconstructed from freehand-tracked 2D B-scans. A qualitative slice guidance method for fine registration is introduced to reduce the placement error caused by target motion. By incorporating the 3D MW specific absorption rate (SAR) model into the heat transfer equation, the MW thermal field simulation module determines the MW power level and the coagulation time for improved ablation therapy. Two types of wrists are developed for the robot: a ,remote centre of motion' (RCM) wrist and a non-RCM wrist, which is preferred in real applications. Results The needle placement accuracies were < 3 mm for both wrists in the mechanical phantom experiment. The target accuracy for the robot with the RCM wrist was improved to 1.6 ± 1.0 mm when real-time 2D US feedback was used in the artificial-tissue phantom experiment. By using the slice guidance method, the robot with the non-RCM wrist achieved accuracy of 1.8 ± 0.9 mm in the ex vivo experiment; even target motion was introduced. In the thermal field experiment, a 5.6% relative mean error was observed between the experimental coagulated neurosis volume and the simulation result. Conclusion The proposed robotic system holds promise to enhance the clinical performance of percutaneous MW ablation of malignant liver tumours. Copyright © 2010 John Wiley & Sons, Ltd. [source] Di(n -butyl) Phthalate Induces Vimentin Filaments Disruption in Rat Sertoli Cells: A Possible Relation with Spermatogenic Cell ApoptosisANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 3 2010M. S. Alam With 3 figures Summary Phthalate esters have been extensively used as a plasticizer of synthetic polymers. Previous studies have revealed that some phthalate esters including di(n -butyl) phthalate (DBP) induce spermatogenic cell apoptosis, although its mechanism is not yet clear. The present study describes that disruption of Sertoli cell vimentin filaments by DBP administration may relate to spermatogenic cell apoptosis. The present histopathological study revealed that a single oral administration of 500 mg/kg DBP caused progressive detachment and displacement of spermatogenic cells away from the seminiferous epithelium and sloughing of them into the lumen. Degenerative spermatogenic cells characterized by chromatin condensation were frequently observed in DBP-treated rats. Ultrastructurally, the degenerative spermatogenic cells were separated from their neighbours, and a collapse of Sertoli cell vimentin filaments was recognized in DBP-treated rats. Sertoli cell cultures showed the increased number and size of vacuoles in their cytoplasm. In agreement with the in vivo experiment, vimentin filaments clearly showed a gradual collapse in DBP-exposed Sertoli cells in vitro. These in vivo and in vitro experiments indicate that DBP-induced collapse of Sertoli cell vimentin filaments may lead to detachment of spermatogenic cells, and then detached cells may undergo apoptosis because of loss of the support and nurture provided by Sertoli cells. [source] Simultaneous Automatic Control of Oxygen and Carbon Dioxide Blood Gases During Cardiopulmonary BypassARTIFICIAL ORGANS, Issue 6 2010Berno J.E. Misgeld Abstract In this work an automatic control strategy is presented for the simultaneous control of oxygen and carbon dioxide blood gas partial pressures to be used during cardiopulmonary bypass surgery with heart,lung machine support. As the exchange of blood gases in the artificial extracorporeal lung is a highly nonlinear process comprising varying time delays, uncertainties, and time-varying parameters, it is currently being controlled manually by specially trained perfusionist staff. The new control strategy includes a feedback linearization routine with augmented time-delay compensation and two external linear gain-scheduled controllers, for partial oxygen and carbon dioxide pressures. The controllers were robustly tuned and tested in simulations with a detailed artificial lung (oxygenator) model in cardiopulmonary bypass conditions. Furthermore, the controllers were implemented in an ex vivo experiment using fresh porcine blood as a substitute fluid and a special deoxygenation technique to simulate a patient undergoing cardiopulmonary bypass. Both controllers showed robust stability during the experiments and a good disturbance rejection to extracorporeal blood flow changes. This automatic control strategy is proposed to improve patient's safety by fast control reference tracking and good disturbance rejection under varying conditions. [source] Using One Rotary Blood Pump to Produce Separate Pulsatile Circulations in the Upper and Lower Halves of the BodyARTIFICIAL ORGANS, Issue 8 2000Takashi Isoyama Abstract: Separate systemic circulations with pulsatile flow were obtained using 1 rotary blood pump as a left ventricular assist device. The outlet of the pump was divided into 2 conduits, 1 connected to the upper half of the body and the other connected to the lower half. An electric actuator that clamped the 2 outlet conduits alternately provided pulsatile flows. An in vitro experiment showed that the pulsatility phases of the upper and lower halves of the body were complementary with pulsatile flow, and an in vivo experiment showed that controlled flow distributions of continuous flows could be obtained. [source] The use of porous calcium phosphate scaffolds with transforming growth factor beta 1 as an onlay bone graft substituteCLINICAL ORAL IMPLANTS RESEARCH, Issue 6 2004An experimental study in rats Abstract Objectives: Autogeneous bone grafting is regarded to be the golden standard for onlay grafts, but it requires a harvesting procedure and the remodeling pattern over time is unpredictable. New materials are constantly being sought to overcome these problems. An in vivo experiment was carried out to evaluate whether (1) porous calcium phosphate cement is a suitable biomaterial for onlay bone grafting, and (2) the addition of transforming growth factor beta 1 (TGF-,1) accelerates de novo bone formation inside the cement porosity. Material and methods: A carrier of porous calcium phosphate cement (Calcibon®) was designed and 16 rats received one preshaped implant each. In 8 out of 16 implants 0.75 ,g TGF-,1 was applied. The animals were killed after 4 weeks and the characteristics of tissue ingrowth into the onlay graft were evaluated. Results: Histologic and quantitative histomorphometrical measurements demonstrated osteoid-like tissue formation in both experimental groups. The addition of TGF-,1 did not induce significantly more osteoid-like tissue formation. On the other hand, in TGF-,-loaded implants, a higher number of pores contained an inflammatory infiltrate. Conclusion: This study indicated that porous calcium phosphate cement is a promising material for clinical situations where bone formation has to be supported. Résumé La greffe osseuse autogčne est considérée comme la meilleure technique actuelle pour les greffons onlay mais elle requiert un processus de prélevement et le remodelage qui s'en suit est imprévisible. De nouveaux matériaux sont donc constamment recherchés. Cette étude in vitro a essayé d'évaluer si 1) le cément phosphate calcium poreux était un biomatériel favorable pour le greffage osseux onlay, 2) si l'addition de TGF-,1 accélérait la néoformation osseuse ŕ l'intérieur de la porosité du cément. Un porteur de cément phosphate calcium poreux (Calcibon®) a été fabriqué et seize rats ont reçu chacun un implant prédécoupé. Au niveau de huit des seize implants 0,75 ,g de TGF ,1 a été appliqué. Les animaux ont été euthanasiés aprčs quatre semaines et les caractéristiques de la croissance interne tissulaire dans le greffon onlay ont étéévaluées. Les mesures histologiques et histomorphométriques quantitatives ont démontré une formation tissulaire semblable ŕ l'ostéogénie dans les deux groupes expérimentaux. L'addition de TGF-ß1 n'induisait pas plus de formation tissulaire ressemblant ŕ celle d'ostéogénie. D'un autre côté, dans les implants chargés de TGF-,1, un nombre plus important de pores contenaient un infiltrat inflammatoire. Cette étude indique que le cément phosphate calcium poreux est un matériau prometteur pour les situations cliniques dans lesquelles la formation osseuse doit ętre améliorée. Zusammenfassung Ziel: Die Transplantation von autologem Knochen wird heute als Goldstandard für die Onlay-Transplantate betrachtet. Es braucht dazu aber einen zusätzlichen Eingriff für die Entnahme und eine Prognose bezüglich der anschliessenden Remodellationsvorgänge sind kaum möglich. Man sucht ständig nach neuen Produkten, um diese Probleme zu überwinden. Man führte eine in vivo Studie durch und untersucht, ob (1) ein poröser Kalziumphosphatzement ein brauchbares Biomaterial für ein Onlay-Transplantat ist, und (2) der Zusatz von TGF-,1 die Neubildung von Knochen in den Porositäten des Zementes positiv beeinflusst. Material und Methode: Man entwickelte einen Trägerzement aus porösem Kalziumphosphat (Calcibon®) und 16 Ratten erhielten je ein vorgeformtes Implantat eingesetzt. Bei 8 der 16 Implantate fügte man zusätzlich 0.75 ,g TGF-,1 dazu. Vier Wochen später opferte man die Tiere und konnte nun die Charakteristika des in die Implantate einwachsenden Gewebes untersuchen. Resultate: Die histologischen und quantitativen histomorphometrischen Messungen zeigten in beiden experimentellen Gruppen osteoidähnliche Gewebsbildungen. Der Zusatz von TGF-,1 bewirkte keine signifikante Zunahme dieser osteoidähnlichen Gewebsbildungen. Die mit TGF-,1 durchsetzten Implantate enthielten aber mehr mit entzündlichem Infiltrat angefüllte Poren. Zusammenfassung: Diese Arbeit zeigte uns, dass ein poröser Kalziumphosphatzement bei klinischen Situationen, wo die Knochenbildung unterstützt werden muss, ein erfolgsversprechendes Material ist. Resumen Objetivos: El injerto de hueso autógeno está considerado como el estándar de oro para injertos superpuestos, pero requiere un procedimiento de recolección y el patrón de remodelado a lo largo del tiempo es impredecible. Constantemente se están buscando materiales nuevos para superar estos problemas. Se llevó a cabo un experimento in vivo para evaluar si (1) el cemento de fosfato cálcico poroso es un biomaterial apropiado para injerto óseo superpuesto, y (2) la adición de TGF-,1 acelera la formación de hueso de novo dentro de la porosidad del cemento. Material y Métodos: Se diseńó un portador de cemento de fosfato cálcico (Calcibon®) y 16 ratas recibieron un implante preformado cada una. En 8 de 16 implantes se aplicaron 0.75 ,g de TGF-,1. Los animales se sacrificaron tras 4 semanas y se evaluaron las características del tejido crecido hacia adentro del injerto superpuesto. Resultados: Las mediciones histológicas e histomorfométricas cuantitativas demostraron formación de tejido tipo osteoide en ambos grupos experimentales. La adición de TGF-,1 no indujo significativamente más formación de tejido tipo osteoide. Por otro lado, en los implantes cargados con TGF-,1, un mayor número de de poros contenían infiltrado inflamatorio. Conclusión: Este estudio indica que el cemento de fosfato cálcico poroso es un material prometedor para situaciones clínicas donde la formación de hueso ha de ser favorecida. [source] A new temperature-sensitive contrast mechanism for MRI: Curie temperature transition-based imagingCONTRAST MEDIA & MOLECULAR IMAGING, Issue 1 2007F. Settecase Abstract A temperature-sensitive MRI contrast mechanism is proposed based on the physical property, the Curie temperature (Tc), at which a ferromagnetic material transitions to paramagnetic state and vice versa. To evaluate the feasibility of this new contrast mechanism, experiments were performed with solid gadolinium metal, which has a Tc of 20°C. In phantom and ex vivo experiments, the magnetic susceptibility artifact area decreased with increasing temperature transitioning across Tc (p,<,0.05). Similar results would be expected for a variety of ferromagnetic substances with substance-specific Tc values. Temperature-sensitive MRI contrast agents harnessing this mechanism may be used to (1) indicate regional attainment of specific temperatures in thermotherapy, (2) render an accumulated contrast agent more or less visible by the external application of appropriate heating or cooling, or (3) quantify tissue temperature based on MR image characteristics and magnetic susceptibility artifact caused by a ferromagnetic,paramagnetic transitioning substance. Copyright © 2007 John Wiley & Sons, Ltd. [source] A diffusible signal attracts olfactory sensory axons toward their target in the developing brain of the mothDEVELOPMENTAL NEUROBIOLOGY, Issue 1 2003Lynne A. Oland Abstract The signals that olfactory receptor axons use to navigate to their target in the CNS are still not well understood. In the moth Manduca sexta, the primary olfactory pathway develops postembryonically, and the receptor axons navigate from an experimentally accessible sensory epithelium to the brain along a pathway long enough for detailed study of regions in which axon behavior changes. The current experiments ask whether diffusible factors contribute to receptor axon guidance. Explants were made from the antennal receptor epithelium and co-cultured in a collagen gel matrix with slices of various regions of the brain. Receptor axons were attracted toward the central regions of the brain, including the protocerebrum and antennal lobe. Receptor axons growing into a slice of the most proximal region of the antennal nerve, where axon sorting normally occurs, showed no directional preference. When the antennal lobe was included in the slice, the receptor axons entering the sorting region grew directly toward the antennal lobe. Taken together with the previous in vivo experiments, the current results suggest that an attractive diffusible factor can serve as one cue to direct misrouted olfactory receptor axons toward the medial regions of the brain, where local cues guide them to the antennal lobe. They also suggest that under normal circumstances, in which the receptor axons follow a pre-existing pupal nerve to the antennal lobe, the diffusible factor emanating from the lobe acts in parallel and at short range to maintain the fidelity of the path into the antennal lobe. © 2003 Wiley Periodicals, Inc. J Neurobiol 56: 24,40, 2003 [source] Comparison of the antilipolytic effects of an A1 adenosine receptor partial agonist in normal and diabetic ratsDIABETES OBESITY & METABOLISM, Issue 2 2009A. K. Dhalla Introduction and Aims:, Elevated plasma free fatty acid (FFA) concentrations play a role in the pathogenesis of type 2 diabetes (2DM). Antilipolytic agents that reduce FFA concentrations may be potentially useful in the treatment of 2DM. Our previous observation that CVT-3619 lowered plasma FFA and triglyceride concentrations in rats and enhanced insulin sensitivity in rodents with dietary-induced forms of insulin resistance suggested that it might be of use in the treatment of patients with 2DM. The present study was undertaken to compare the antilipolytic effects of CVT-3619 in normal (Sprague Dawley, SD) and Zucker diabetic fatty (ZDF) rats. Results:, ZDF rats had significantly higher fat pad weight, glucose, insulin and FFA concentrations than those of SD rats. EC50 values for forskolin-stimulated FFA release from isolated adipocytes from SD and ZDF rats were 750 and 53 nM, respectively (p < 0.05). Maximal forskolin stimulation of FFA release was significantly (p < 0.01) less in ZDF rats (133 ± 60 ,M) compared with SD rats (332 ± 38 ,M). EC50 values for isoproterenol to increase lipolysis in adipocytes from SD and ZDF rats were 2 and 7 nM respectively. Maximal isoproterenol-stimulated lipolysis was significantly (p < 0.01) lower in adipocytes from ZDF rats (179 ± 23 ,M) compared with SD rats (343 ± 27 ,M). Insulin inhibited lipolysis in adipocytes from SD rats with an IC50 value of 30 pM, whereas adipocytes from ZDF rats were resistant to the antilipolytic actions of insulin. In contrast, IC50 values for CVT-3619 to inhibit the release of FFA from SD and ZDF adipocytes were essentially the same (63 and 123 nM respectively). CVT-3619 inhibited lipolysis more than insulin in both SD (86 vs. 46%, p < 0.001) and ZDF (80 vs. 13%, p < 0.001) adipocytes. In in vivo experiments, CVT-3619 (5 mg/kg, PO) lowered FFA to a similar extent in both groups. Plasma concentrations of CVT-3619 were not different in SD and ZDF rats. There was no significant difference in the messenger RNA expression of the A1 receptors relative to ,-actin expression in adipocytes from SD (0.98 ± 0.2) and ZDF rats (0.99 ± 0.3). Conclusion:, The antilipolytic effects of CVT-3619 appear to be independent of insulin resistance and animal model. [source] Intravascular neural interface with nanowire electrodeELECTRONICS & COMMUNICATIONS IN JAPAN, Issue 7 2009Hirobumi Watanabe Abstract A minimally invasive electrical recording and stimulating technique capable of simultaneously monitoring the activity of a significant number (e.g., 103 to 104) of neurons is an absolute prerequisite in developing an effective brain,machine interface. Although there are many excellent methodologies for recording single or multiple neurons, there has been no methodology for accessing large numbers of cells in a behaving experimental animal or human individual. Brain vascular parenchyma is a promising candidate for addressing this problem. It has been proposed [1, 2] that a multitude of nanowire electrodes introduced into the central nervous system through the vascular system to address any brain area may be a possible solution. In this study we implement a design for such microcatheter for ex vivo experiments. Using Wollaston platinum wire, we design a submicron-scale electrode and develop a fabrication method. We then evaluate the mechanical properties of the electrode in a flow when passing through the intricacies of the capillary bed in ex vivo Xenopus laevis experiments. Furthermore, we demonstrate the feasibility of intravascular recording in the spinal cord of Xenopus laevis. © 2009 Wiley Periodicals, Inc. Electron Comm Jpn, 92(7): 29,37, 2009; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ecj.10058 [source] Ecdysteroid synthesis and imaginal disc development in the midge Chironomus riparius as biomarkers for endocrine effects of tributyltinENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 5 2002Torsten Hahn Abstract Acute effects of the endocrine disruptor bis (tri- n -butyltin) oxide (TBTO) on molting-hormone biosynthesis and imaginaldisc development were investigated in larvae of the midge Chironomus riparius (Meigen). Ecdysteroid synthesis was measured by 24-h incubation of molting-hormone-synthesizing tissues (prothoracic glands) in vitro with or without the addition of TBTO. The amount of ecdysteroids produced was analyzed by radioimmunoassay. Developmental effects in vivo were investigated by determining the developmental phase of the genital imaginal discs before and after a 48-h exposure to TBTO in water. Sex-specific effects were found with both endpoints. Ecdysteroid synthesis was significantly reduced (analysis of variance [ANOVA], p , 0.005) in female larvae at all concentrations (TBTO-Sn at 50, 500, and 5,000 ng/L), whereas a significant elevation of the biosynthesis rate occurred in male larvae in the 500-ng/L treatment (ANOVA, p , 0.05). In vivo experiments with development of the genital imaginal disc within a 48-h exposure period revealed a significantly slower development in female larvae and a significantly faster development in male larvae (contingency tables, p , 0.001) at all concentrations tested (TBTO-Sn at 10, 50, 200, and 1,000 ng/L). These results partly coincided with the in vitro effects on molting-hormone synthesis. The 48-h median lethal concentration (LC50) was 25 ,g/L (20,30 ,g/L 95% confidence intervals). The combination of in vitro and in vivo methods has proven to be a useful approach for the detection of endocrine effects of TBTO in C. riparius at levels 2,000-fold below the LC50 value. High sensitivity and short test duration suggest that chironomids may have potential as freshwater sentinel organisms for endocrine-disrupting chemicals. [source] Development of equine upper airway fluid mechanics model for Thoroughbred racehorsesEQUINE VETERINARY JOURNAL, Issue 3 2008V. RAKESH Summary Reason for performing study: Computational fluid dynamics (CFD) models provide the means to evaluate airflow in the upper airways without requiring in vivo experiments. Hypothesis: The physiological conditions of a Thoroughbred racehorse's upper airway during exercise could be simulated. Methods: Computed tomography scanned images of a 3-year-old intact male Thoroughbred racehorse cadaver were used to simulate in vivo geometry. Airway pressure traces from a live Thoroughbred horse, during exercise was used to set the boundary condition. Fluid-flow equations were solved for turbulent flow in the airway during inspiratory and expiratory phases. The wall pressure turbulent kinetic energy and velocity distributions were studied at different cross-sections along the airway. This provided insight into the general flow pattern and helped identify regions susceptible to dynamic collapse. Results: The airflow velocity and static tracheal pressure were comparable to data of horses exercising on a high-speed treadmill reported in recent literature. The cross-sectional area of the fully dilated rima glottidis was 7% greater than the trachea. During inspiration, the area of highest turbulence (i.e. kinetic energy) was in the larynx, the rostral aspect of the nasopharynx was subjected to the most negative wall pressure and the highest airflow velocity is more caudal on the ventral aspect of the nasopharynx (i.e. the soft palate). During exhalation, the area of highest turbulence was in the rostral and mid-nasopharynx, the maximum positive pressure was observed at the caudal aspect of the soft palate and the highest airflow velocity at the front of the nasopharynx. Conclusions and clinical relevance: In the equine upper airway collapsible area, the floor of the rostral aspect of the nasopharynx is subjected to the most significant collapsing pressure with high average turbulent kinetic during inhalation, which may lead to palatal instability and explain the high prevalence of dorsal displacement of the soft palate (DDSP) in racehorses. Maximal abduction of the arytenoid cartilage may not be needed for optimal performance, since the trachea cross-sectional area is 7% smaller than the rima glottidis. [source] Deep brain stimulation mechanisms: beyond the concept of local functional inhibitionEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 7 2010Jean-Michel Deniau Abstract Deep brain electrical stimulation has become a recognized therapy in the treatment of a variety of motor disorders and has potentially promising applications in a wide range of neurological diseases including neuropsychiatry. Behavioural observation that electrical high-frequency stimulation of a given brain area induces an effect similar to a lesion suggested a mechanism of functional inhibition. In vitro and in vivo experiments as well as per operative recordings in patients have revealed a variety of effects involving local changes of neuronal excitability as well as widespread effects throughout the connected network resulting from activation of axons, including antidromic activation. Here we review current data regarding the local and network activity changes induced by high-frequency stimulation of the subthalamic nucleus and discuss this in the context of motor restoration in Parkinson's disease. Stressing the important functional consequences of axonal activation in deep brain stimulation mechanisms, we highlight the importance of developing anatomical knowledge concerning the fibre connections of the putative therapeutic targets. [source] The murine neurokinin NK1 receptor gene contributes to the adult hypoxic facilitation of ventilationEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 12 2002Krzysztof Ptak Abstract Substance P and neurokinin-1 receptors (NK1) modulate the respiratory activity and are expressed early during development. We tested the hypothesis that NK1 receptors are involved in prenatal development of the respiratory network by comparing the resting respiratory activity and the respiratory response to hypoxia of control mice and mutant mice lacking the NK1 receptor (NK1,/,). In vitro and in vivo experiments were conducted on neonatal, young and adult mice from wild-type and NK1,/, strains. In the wild strain, immunohistological, pharmacological and electrophysiological studies showed that NK1 receptors were expressed within medullary respiratory areas prior to birth and that their activation at birth modulated central respiratory activity and the membrane properties of phrenic motoneurons. Both the membrane properties of phrenic motoneurons and the respiratory activity generated in vitro by brainstem-spinal cord preparation from NK1,/, neonate mice were similar to that from the wild strain. In addition, in vivo ventilation recordings by plethysmography did not reveal interstrain differences in resting breathing parameters. The facilitation of ventilation by short-lasting hypoxia was similar in wild and NK1,/, neonates but was significantly weaker in adult NK1,/, mice. Results demonstrate that NK1 receptors do appear to be necessary for a normal respiratory response to short-lasting hypoxia in the adult. However, NK1 receptors are not obligatory for the prenatal development of the respiratory network, for the production of the rhythm, or for the regulation of breathing by short-lasting hypoxia in neonates. [source] 17,-estradiol induces aromatase activity in intact human anagen hair follicles ex vivoEXPERIMENTAL DERMATOLOGY, Issue 4 2002R. Hoffmann Abstract: For topical treatment of androgenetic alopecia (AGA) in women, solutions containing either estradiol benzoate, estradiol valerate, 17,- or 17,-estradiol are commercially available in Europe and some studies show an increased anagen and decreased telogen rate after treatment as compared with placebo. At present it is not precisely known how estrogens mediate their beneficial effect on AGA-affected hair follicles. We have shown recently that 17,-estradiol is able to diminish the amount of dihydrotestosterone (DHT) formed by human hair follicles after incubation with testosterone, while increasing the concentration of weaker steroids such as estrogens. Because aromatase is involved in the conversion of testosterone to estrogens and because there is some clinical evidence that aromatase activity may be involved in the pathogenesis of AGA, we addressed the question whether aromatase is expressed in human hair follicles and whether 17,-estradiol is able to modify the aromatase activity. Herewith we were able to demonstrate that intact, microdissected hair follicles from female donors express considerably more aromatase activity than hair follicles from male donors. Using immunohistochemistry, we detected the aromatase mainly in the epithelial parts of the hair follicle and not in the dermal papilla. Furthermore, we show that in comparison to the controls, we noticed in 17,-estradiol-incubated (1 nM) female hair follicles a concentration- and time-dependent increase of aromatase activity (at 24 h: 1 nM = +18%, 100 nM = +25%, 1 µM = +57%; 24 h: 1 nM = +18%, 48 h: 1 nM = +25%). In conclusion, our ex vivo experiments suggest that under the influence of 17,-estradiol an increased conversion of testosterone to 17,-estradiol and androstendione to estrone takes place, which might explain the beneficial effects of estrogen treatment of AGA. [source] Leishmania infantum LeIF protein is an ATP-dependent RNA helicase and an eIF4A-like factor that inhibits translation in yeastFEBS JOURNAL, Issue 22 2006Mourad Barhoumi LeIF, a Leishmania protein similar to the eukaryotic initiation factor eIF4A, which is a prototype of the DEAD box protein family, was originally described as a Th1-type natural adjuvant and as an antigen that induces an IL12-mediated Th1 response in the peripheral blood mononuclear cells of leishmaniasis patients. This study aims to characterize this protein by comparative biochemical and genetic analysis with eIF4A in order to assess its potential as a target for drug development. We show that a His-tagged, recombinant, LeIF protein of Leishmania infantum, which was purified from Escherichia coli, is both an RNA-dependent ATPase and an ATP-dependent RNA helicase in vitro, as described previously for other members of the DEAD box helicase protein family. In vivo experiments show that the LeIF gene cannot complement the deletion of the essential TIF1 and TIF2 genes in the yeast Saccharomyces cerevisiae that encode eIF4A. In contrast, expression of LeIF inhibits yeast growth when endogenous eIF4A is expressed off only one of its two encoding genes. Furthermore, in vitro binding assays show that LeIF interacts with yeast eIF4G. These results show an unproductive interaction of LeIF with translation initiation factors in yeast. Furthermore, the 25 amino terminal residues were shown to enhance the ability of LeIF to interfere with the translation machinery in yeast. [source] The effect of thiamine supplementation on tumour proliferationFEBS JOURNAL, Issue 15 2001A metabolic control analysis study Thiamine deficiency frequently occurs in patients with advanced cancer and therefore thiamine supplementation is used as nutritional support. Thiamine (vitamin B1) is metabolized to thiamine pyrophosphate, the cofactor of transketolase, which is involved in ribose synthesis, necessary for cell replication. Thus, it is important to determine whether the benefits of thiamine supplementation outweigh the risks of tumor proliferation. Using oxythiamine (an irreversible inhibitor of transketolase) and metabolic control analysis (MCA) methods, we measured an in vivo tumour growth control coefficient of 0.9 for the thiamine-transketolase complex in mice with Ehrlich's ascites tumour. Thus, transketolase enzyme and thiamine clearly determine cell proliferation in the Ehrlich's ascites tumour model. This high control coefficient allows us to predict that in advanced tumours, which are commonly thiamine deficient, supplementation of thiamine could significantly increase tumour growth through transketolase activation. The effect of thiamine supplementation on tumour proliferation was demonstrated by in vivo experiments in mice with the ascites tumour. Thiamine supplementation in doses between 12.5 and 250 times the recommended dietary allowance (RDA) for mice were administered starting on day four of tumour inoculation. We observed a high stimulatory effect on tumour growth of 164% compared to controls at a thiamine dose of 25 times the RDA. This growth stimulatory effect was predicted on the basis of correction of the pre-existing level of thiamine deficiency (42%), as assayed by the cofactor/enzyme ratio. Interestingly, at very high overdoses of thiamine, ,,2500 times the RDA, thiamine supplementation had the opposite effect and caused 10% inhibition of tumour growth. This effect was heightened, resulting in a 36% decrease, when thiamine supplementation was administered from the 7th day prior to tumour inoculation. Our results show that thiamine supplementation sufficient to correct existing thiamine deficiency stimulates tumour proliferation as predicted by MCA. The tumour inhibitory effect at high doses of thiamine is unexplained and merits further study. [source] Impaired synthesis and secretion of SopA in Salmonella Typhimurium dam mutantsFEMS MICROBIOLOGY LETTERS, Issue 1 2009Mónica N. Giacomodonato Abstract DNA adenine methylation regulates virulence gene expression in certain bacteria, including Salmonella Typhimurium. The aim of this study was to investigate the involvement of DNA adenine methylase (Dam) methylation in the expression and secretion of the SPI-1 effector protein SopA. For this purpose, SopA,FLAG-tagged wild-type and dam strains of Salmonella Typhimurium were constructed. The expression and secretion of SopA were determined in bacterial culture and in intracellular bacteria recovered from infected HEp-2 epithelial cells. Bacterial culture supernatants and pellets were used to investigate secreted proteins and cell-associated proteins, respectively. Western blot and quantitative reverse transcriptase PCR analysis showed that the dam mutant expresses lower levels of SopA than the wild-type strain. Interestingly, the strain lacking Dam synthesizes SopA under nonpermissive conditions (28 °C). In addition, SopA secretion was drastically impaired in the dam mutant. In vivo experiments showed that the intracellular Salmonella dam mutant synthesizes SopA although in lower amounts than the wild-type strain. Taken together, our results suggest that Dam methylation modulates the expression and secretion of SopA in Salmonella Typhimurium. [source] The aerodynamics and efficiency of wind pollination in grassesFUNCTIONAL ECOLOGY, Issue 4 2010James E. Cresswell Summary 1.,Under natural selection for sexual success, the reproductive organs of plants should evolve to become highly effective pollen receptors. Among wind-pollinated plants, larger reproductive structures appear counter-adapted to accumulate pollen by impaction on their windward surfaces, because airborne particles are less able to penetrate the thicker boundary layer of larger targets. Therefore, it has been proposed that wind-pollinated plants with pollen receptors on relatively large structures, like some grasses (family Poaceae), are architecturally adapted to create downstream vortices in which airborne pollen recirculates before accumulating on leeward surfaces. From this basis, the striking diversity among the grasses in the architecture of their flowering stems has been attributed in part to the existence of these contrasting mechanisms for effecting pollen receipt, namely impact collection and recirculatory collection. 2.,We investigated the relative importance of impact and recirculatory collection in grasses by analysing a model system in silico using Computational Fluid Dynamics and by conducting in vivo experiments, both in a wind tunnel and outdoors, using two grass species with compact inflorescences, Alopecurus pratensis and Anthoxanthum odoratum. 3.,Irrespective of the experimental approach, we found that although pollen recirculated in the leeward eddies of inflorescences, over 95% of the accumulated pollen was collected by windward surfaces. 4.,In A. pratensis, the collection efficiency (proportion of oncoming pollen collected) was between 5% and 20%, depending on wind speed in the range 0·5,1·9 m s,1 and these levels conform to those predicted by a mechanistic model of impact collection. 5.,Our results demonstrate that grass species with larger inflorescences are, like those with smaller inflorescences, primarily impact collectors of airborne pollen, which suggests that dissimilar reproductive morphology among species cannot be attributed to differentiation in the mode of pollen capture and, instead, requires reference to other factors, such as the need to produce, protect and disperse seeds of different sizes in different environments. [source] Hypericum caprifoliatum (Guttiferae) Cham.FUNDAMENTAL & CLINICAL PHARMACOLOGY, Issue 6 2006& Schltdl.: a species native to South Brazil with antidepressant-like activity Abstract In this work, previously published and unpublished results on biological activity of Hypericum caprifoliatum, a native specie to South Brazil, are presented. Lipophilic extracts obtained from this species showed an antidepressant-like activity in mice and rat forced swimming test. Results from in vivo experiments suggest an effect on the dopaminergic transmission. Besides that, in vitro experiments demonstrated that the extract and its main component (a phloroglucinol derivative) inhibit monoamine uptake in a concentration-dependent manner, more potently to dopamine, but this effect is not related to direct binding at the uptake sites. It was also observed that a 3-day treatment with lipophilic extract prevents stress-induced corticosterone rise in mice frontal cortex but not in plasma. The lipophilic and methanolic H. caprifoliatum extracts also demonstrated antinociceptive effect, which seems to be indirectly mediated by the opioid system. These results indicate that H. caprifoliatum presents a promising antidepressant-like effect in rodents which seems to be related to a mechanism different from that of other classes of antidepressants. [source] Involvement of the MP1 scaffold protein in ERK signaling regulation during Drosophila wing developmentGENES TO CELLS, Issue 11 2008Emmanučle Mouchel-Vielh Mitogen-activated protein kinase (MAPK) cascades are evolutionary conserved transduction pathways involved in many cellular processes. Kinase modules are associated with scaffold proteins that regulate signaling by providing critical spatial and temporal specificities. Some of these scaffold proteins have been shown to be conserved, both in sequence and function. In mouse, the scaffold MP1 (MEK Partner 1) forms a signaling complex with MEK1 and ERK1. In this work, we focus on Drosophila MP1 (dMP1). We show that dMP1 is expressed ubiquitously during embryonic and larval development. By in vitro and in vivo experiments, we show that dMP1 is located in the cytoplasm and the nuclei, and that it interacts with MEK and ERK. Genetic studies with transgenic Drosophila lines allowing either dMP1 over-expression or dMP1 down-regulation by RNA interference highlight dMP1 function in the control of cell differentiation during development of the Drosophila wing. [source] An MLCK-dependent window in late G1 controls S phase entry of proliferating rodent hepatocytes via ERK-p70S6K pathway,HEPATOLOGY, Issue 1 2006Anne Bessard We show that MLCK (myosin light chain kinase) plays a key role in cell cycle progression of hepatocytes: either chemical inhibitor ML7 or RNA interference led to blockade of cyclin D1 expression and DNA replication, providing evidence that MLCK regulated S phase entry. Conversely, inhibition of RhoK by specific inhibitor Y27632 or RhoK dominant-negative vector did not influence progression in late G1 and S phase entry. Inhibition of either MLCK or RhoK did not block ERK1/2 phosphorylation, whereas MLCK regulated ERK2-dependent p70S6K activation. In addition, DNA synthesis was reduced in hepatocytes treated with p70S6K siRNA, demonstrating the key role played by the kinase in S phase entry. Interestingly, after the G1/S transition, DNA replication in S phase was no longer dependent on MLCK activity. We strengthened this result by ex vivo experiments and evidenced an MLCK-dependent window in late G1 phase of regenerating liver after two-thirds partial hepatectomy. In conclusion, our results underline an MLCK-dependent restriction point in G1/S transition, occurring downstream of ERK2 through the regulation of p70S6K activation, and highlighting a new signaling pathway critical for hepatocyte proliferation. (HEPATOLOGY 2006;44:152,163.) [source] Noncontact measurement of deep tissue absorption coefficient using Spatially Resolved Near-Infrared SpectroscopyIEEJ TRANSACTIONS ON ELECTRICAL AND ELECTRONIC ENGINEERING, Issue 4 2007Masatsugu Niwayama Member Abstract We examined the influence of probe-tissue distance on the relationship between the deep tissue absorption coefficient and the spatial profile of light intensity by Monte Carlo simulation and in vivo experiments, using noncontact spatially resolved near-infrared spectroscopy (SRS). These results suggest that noncontact SRS can be reliably used for noncontact measurement of the absorption coefficient and oxygen saturation of deep tissues. © 2007 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc. [source] Mesenchymal stem cells enhance growth and metastasis of colon cancerINTERNATIONAL JOURNAL OF CANCER, Issue 10 2010Kei Shinagawa Abstract Recently, mesenchymal stem cells (MSCs) were reported to migrate to tumor stroma as well as injured tissue. We examined the role of human MSCs in tumor stroma using an orthotopic nude mice model of KM12SM colon cancer. In in vivo experiments, systemically injected MSCs migrated to the stroma of orthotopic colon tumors and metastatic liver tumors. Orthotopic transplantation of KM12SM cells mixed with MSCs resulted in greater tumor weight than did transplantation of KM12SM cells alone. The survival rate was significantly lower in the mixed-cell group, and liver metastasis was seen only in this group. Moreover, tumors resulting from transplantation of mixed cells had a significantly higher proliferating cell nuclear antigen labeling index, significantly greater microvessel area and significantly lower apoptotic index. Splenic injection of KM12SM cells mixed with MSCs, in comparison to splenic injection of KM12SM cells alone, resulted in a significantly greater number of liver metastases. MSCs incorporated into the stroma of primary and metastatic tumors expressed ,-smooth muscle actin and platelet-derived growth factor receptor-, as carcinoma-associated fibroblast (CAF) markers. In in vitro experiments, KM12SM cells recruited MSCs, and MSCs stimulated migration and invasion of tumor cells through the release of soluble factors. Collectively, MSCs migrate and differentiate into CAFs in tumor stroma, and they promote growth and metastasis of colon cancer by enhancing angiogenesis, migration and invasion and by inhibiting apoptosis of tumor cells. [source] Kojic acid reduces the cytotoxic effects of sulfur mustard on cultures containing human melanoma cells in vitroJOURNAL OF APPLIED TOXICOLOGY, Issue 6 2001C. N. Smith Abstract In vivo experiments have shown that melanocytes are more sensitive than keratinocytes to the cytotoxic effects of sulfur mustard when it is applied topically to pig skin.1 It has been hypothesized that this is caused by the uncoupling of the melanogenic pathway by depletion of cellular glutathione, resulting in the uncontrolled production of cytotoxic quinone free-radical species by tyrosinase.2. In the present study, the feasibility of blocking the melanogenic pathway as a means of reducing the cytotoxicity of sulfur mustard was evaluated using kojic acid. Kojic acid is a topically applied depigmenting agent that exerts its effect by acting as a slow-binding, competitive inhibitor of tyrosinase.3 Preincubation of G361 pigmented melanoma cells and mixed cultures of G361 cells and SVK keratinocytes with 2.5 mM kojic acid resulted in significant increases in the viability of these cultures as determined by neutral red (NR) and gentian violet (GV) dye binding assays for up to 48 h following exposure to 50 µM sulfur mustard. The highest levels of protection were seen in the G361 cultures, with a 26.8% increase in culture viability (NR assay) compared with the sulfur-mustard-only controls at 24 h. Preincubation of SVK cells alone with kojic acid resulted in lower increases in viability (2.5% at 24 h by the NR assay). Inhibition of the melanogenic pathway reduces the sensitivity of cultures containing pigment cells to sulfur mustard. © Crown copyright 2001. Reproduced with the permission of Her Majesty's Stationery Office. 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