Home About us Contact
|
Home About us Contact | ||
|
|
||
Different Protocols (different + protocol)
Selected AbstractsBismuth/Polyaniline/Glassy Carbon Electrodes Prepared with Different Protocols for Stripping Voltammetric Determination of Trace Cd and Pb in Solutions Having SurfactantsELECTROANALYSIS, Issue 2 2010M. Wang Abstract To improve reproducibility, stability and sensitivity, a bismuth (Bi) thin film was coated on glassy carbon (GC) substrates which surfaces were modified with a porous thin layer of polyaniline (PANI) via multipulse potentiostatic electropolymerization to form Bi/PANI/GC electrodes (Bi/PANI/GCEs). The Bi/PANI/GCEs were used successfully for simultaneous detection and determination of Cd2+ and Pb2+ ions, and various parameters were studied with reference to square wave anodic stripping voltammetric (SWASV) signals. The experimental results depicted that the environment-friendly Bi/PANI/GCEs had the ability to rapidly monitor trace heavy metals even in the presence of surface-active compounds. [source] Rapid and easy semi-quantitative evaluation method for diacylglycerol and inositol-1,4,5-trisphosphate generation in orexin receptor signallingACTA PHYSIOLOGICA, Issue 3 2010M. E. Ekholm Abstract Aim:, Fluorescent protein-based indicators have enabled measurement of intracellular signals previously nearly inaccessible for studies. However, indicators showing intracellular translocation upon response suffer from serious limitations, especially the very time-consuming data collection. We therefore set out in this study to evaluate whether fixing and counting cells showing translocation could mend this issue. Methods:, Altogether three different genetically encoded indicators for diacylglycerol and inositol-1,4,5-trisphosphate were transiently expressed in Chinese hamster ovary cells stably expressing human OX1 orexin receptors. Upon stimulation with orexin-A, the cells were fixed with six different protocols. Results:, Different protocols showed clear differences in their ability to preserve the indicator's localization (i.e. translocation after stimulus) and its fluorescence, and the best results for each indicator were obtained with a different protocol. The concentration,response data obtained with cell counting are mostly comparable to the real-time translocation and biochemical data. Conclusion:, The counting method, as used here, works at single time point and looses the single-cell-quantitative aspect. However, it also has some useful properties. First, it easily allows processing of a 100- to 1000-fold higher cell numbers than real-time imaging producing statistically consistent population-quantitative data much faster. Secondly, it does not require expensive real-time imaging equipment. Fluorescence in fixed cells can also be quantitated, though this analysis would be more time-consuming than cell counting. Thirdly, in addition to the quantitative data collection, the method could be applied for identifying responsive cells. This might be very useful in identification of e.g. orexin-responding neurones in a large population of non-responsive cells in primary cultures. [source] Proteomic analysis of rat brain tissue: Comparison of protocols for two-dimensional gel electrophoresis analysis based on different solubilizing agentsELECTROPHORESIS, Issue 24 2002Lucia Carboni Abstract The present study reports a comparison of recently described solubilizing methods, to set up a simple protocol for obtaining two-dimensional (2-D) gel electrophoresis maps of brain tissue. Different protocols were used for preparing rat brain homogenates and the resulting maps were compared by image analysis. Three different detergents, two delipidation methods, and introduction of a fractionation step based on different protein solubility in surfactants, were evaluated. When using efficient zwitterionic detergents (3-[(3-cholamidopropyl)dimethylamino]-1-propanesulfonate, CHAPS; amidosulfobetaine 14, ASB-14), the patterns obtained by direct loading of total extracts were qualitatively overlapping with patterns obtained from fractionated samples. In contrast, a weaker nonionic agent (Nonidet P-40, NP-40) produced a different protein pattern in the collected fractions. Delipidation did not improve the results for all the different extraction methods. Immunoblots performed with antibodies recognizing cytosolic and membrane-spanning proteins, which were detected as nondegraded spots, showed that membrane proteins with intermediate molecular mass could be recovered. We suggest, as a simple and efficient method for preparing rat brain maps, the homogenization in a solution containing an efficient zwitterionic surfactant, which allows to solubilize cytosolic and membrane proteins in a single step. Alternatively, a fractionation can be carried out on samples homogenized by a weak solubilizing agent, a more labor-intensive effort resulting in a larger number of proteins on two maps. [source] Lasers for facial rejuvenation: a reviewINTERNATIONAL JOURNAL OF DERMATOLOGY, Issue 6 2003Evangelia Papadavid MD Background Different types of laser are used for resurfacing and collagen remodeling in cutaneous laser surgery. Methods A systematic review was performed of the different types of laser currently employed for skin rejuvenation. These systems are either ablative [high-energy pulsed or scanned carbon dioxide (CO2) laser emitting at a wavelength of 10,600 nm, single- or variable-pulse or dual ablative/coagulative mode erbium:yttrium aluminum garnet (Er:YAG) laser emitting at a wavelength of 2940 nm, or systems combining both 10,600 nm and 2940 nm wavelengths] or nonablative [Q-switched neodymium:yttrium aluminum garnet (Nd:YAG) laser emitting at a wavelength of 1064 nm, Nd:YAG laser emitting at a wavelength of 1320 nm, or diode laser emitting at a wavelength of 1450 nm]. Different protocols, patient selection, treatment techniques, and complications are discussed for each system. Results New-generation CO2 resurfacing lasers have been successful in the treatment of photodamaged skin and scarring, with a postoperative morbidity dependent on the depth of thermal damage. Because of its minimal penetration, the pulsed Er:YAG laser, usually used in the treatment of more superficial rhytides, produces less postoperative morbidity. Novel ablative systems have been developed and a further understanding of laser,tissue interaction has led to the design of nonablative systems for the treatment of rhytides, scarring, and photodamaged skin, the efficacy and profile of which remain to be evaluated in the long term. Conclusions There are several effective techniques for scar revision and the treatment of aged skin, but all have their drawbacks due to a lack of precise depth control and unwanted damage to the lower layers of the dermis. The Er:YAG laser is the treatment of choice for fine lines and superficial scars, whereas the CO2 laser is better for deeper rhytides and scars. In the future, a combination of lasers may be used for facial rejuvenation. [source] Outer membrane proteome of Actinobacillus pleuropneumoniae: LC-MS/MS analyses validate in silico predictionsPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 11 2007Jacqueline W. Chung Abstract The Gram-negative bacterial pathogen Actinobacillus pleuropneumoniae causes porcine pneumonia, a highly infectious respiratory disease that contributes to major economic losses in the swine industry. Outer membrane (OM) proteins play key roles in infection and may be targets for drug and vaccine research. Exploiting the genome sequence of A. pleuropneumoniae serotype 5b, we scanned in silico for proteins predicted to be localized at the cell surface. Five genome scanning programs (Proteome Analyst, PSORT-b, BOMP, Lipo, and LipoP) were run to construct a consensus prediction list of 93 OM proteins in A. pleuropneumoniae. An inventory of predicted OM proteins was complemented by proteomic analyses utilizing gel- and solution-based methods, both coupled to LC-MS/MS. Different protocols were explored to enrich for OM proteins; the most rewarding required sucrose gradient centrifugation followed by membrane washes with sodium bromide and sodium carbonate. This protocol facilitated our identification of 47 OM proteins that represent 50% of the predicted OM proteome, most of which have not been characterized. Our study establishes the first OM proteome of A. pleuropneumoniae. [source] Rapid and easy semi-quantitative evaluation method for diacylglycerol and inositol-1,4,5-trisphosphate generation in orexin receptor signallingACTA PHYSIOLOGICA, Issue 3 2010M. E. Ekholm Abstract Aim:, Fluorescent protein-based indicators have enabled measurement of intracellular signals previously nearly inaccessible for studies. However, indicators showing intracellular translocation upon response suffer from serious limitations, especially the very time-consuming data collection. We therefore set out in this study to evaluate whether fixing and counting cells showing translocation could mend this issue. Methods:, Altogether three different genetically encoded indicators for diacylglycerol and inositol-1,4,5-trisphosphate were transiently expressed in Chinese hamster ovary cells stably expressing human OX1 orexin receptors. Upon stimulation with orexin-A, the cells were fixed with six different protocols. Results:, Different protocols showed clear differences in their ability to preserve the indicator's localization (i.e. translocation after stimulus) and its fluorescence, and the best results for each indicator were obtained with a different protocol. The concentration,response data obtained with cell counting are mostly comparable to the real-time translocation and biochemical data. Conclusion:, The counting method, as used here, works at single time point and looses the single-cell-quantitative aspect. However, it also has some useful properties. First, it easily allows processing of a 100- to 1000-fold higher cell numbers than real-time imaging producing statistically consistent population-quantitative data much faster. Secondly, it does not require expensive real-time imaging equipment. Fluorescence in fixed cells can also be quantitated, though this analysis would be more time-consuming than cell counting. Thirdly, in addition to the quantitative data collection, the method could be applied for identifying responsive cells. This might be very useful in identification of e.g. orexin-responding neurones in a large population of non-responsive cells in primary cultures. [source] Elements of a neurobiological theory of hippocampal function: the role of synaptic plasticity, synaptic tagging and schemasEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 11 2006R. G. M. MorrisArticle first published online: 8 JUN 200 Abstract The 2004 EJN Lecture was an attempt to lay out further aspects of a developing neurobiological theory of hippocampal function [Morris, R.G.M., Moser, E.I., Riedel, G., Martin, S.J., Sandin, J., Day, M. & O'Carroll, C. (2003) Phil. Trans. R. Soc. Lond. B Biol. Sci., 358, 773,786.] These are that (i) activity-dependent synaptic plasticity plays a key role in the automatic encoding and initial storage of attended experience; (ii) the persistence of hippocampal synaptic potentiation over time can be influenced by other independent neural events happening closely in time, an idea with behavioural implications for memory; and (iii) that systems-level consolidation of memory traces within neocortex is guided both by hippocampal traces that have been subject to cellular consolidation and by the presence of organized schema in neocortex into which relevant newly encoded information might be stored. Hippocampal memory is associative and, to study it more effectively than with previous paradigms, a new learning task is described which is unusual in requiring the incidental encoding of flavour,place paired associates, with the readout of successful storage being successful recall of a place given the flavour with which it was paired. NMDA receptor-dependent synaptic plasticity is shown to be critical for the encoding and intermediate storage of memory traces in this task, while AMPA receptor-mediated fast synaptic transmission is necessary for memory retrieval. Typically, these rapidly encoded traces decay quite rapidly over time. Synaptic potentiation also decays rapidly, but can be rendered more persistent by a process of cellular consolidation in which synaptic tagging and capture play a key part in determining whether or not it will be persistent. Synaptic tags set at the time of an event, even many trivial events, can capture the products of the synthesis of plasticity proteins set in train by events before, during or even after an event to be remembered. Tag,protein interactions stabilize synaptic potentiation and, by implication, memory. The behavioural implications of tagging are explored. Finally, using a different protocol for flavour,place paired associate learning, it is shown that rats can develop a spatial schema which represents the relative locations of several different flavours of food hidden at places within a familiar space. This schema is learned gradually but, once acquired, enables new paired associates to be encoded and stored in one trial. Their incorporation into the schema prevents rapid forgetting and suggests that schema play a key and hitherto unappreciated role in systems-level memory consolidation. The elements of what may eventually mature into a more formal neurobiological theory of hippocampal memory are laid out as specific propositions with detailed conceptual discussion and reference to recent data. [source] Rapid and easy semi-quantitative evaluation method for diacylglycerol and inositol-1,4,5-trisphosphate generation in orexin receptor signallingACTA PHYSIOLOGICA, Issue 3 2010M. E. Ekholm Abstract Aim:, Fluorescent protein-based indicators have enabled measurement of intracellular signals previously nearly inaccessible for studies. However, indicators showing intracellular translocation upon response suffer from serious limitations, especially the very time-consuming data collection. We therefore set out in this study to evaluate whether fixing and counting cells showing translocation could mend this issue. Methods:, Altogether three different genetically encoded indicators for diacylglycerol and inositol-1,4,5-trisphosphate were transiently expressed in Chinese hamster ovary cells stably expressing human OX1 orexin receptors. Upon stimulation with orexin-A, the cells were fixed with six different protocols. Results:, Different protocols showed clear differences in their ability to preserve the indicator's localization (i.e. translocation after stimulus) and its fluorescence, and the best results for each indicator were obtained with a different protocol. The concentration,response data obtained with cell counting are mostly comparable to the real-time translocation and biochemical data. Conclusion:, The counting method, as used here, works at single time point and looses the single-cell-quantitative aspect. However, it also has some useful properties. First, it easily allows processing of a 100- to 1000-fold higher cell numbers than real-time imaging producing statistically consistent population-quantitative data much faster. Secondly, it does not require expensive real-time imaging equipment. Fluorescence in fixed cells can also be quantitated, though this analysis would be more time-consuming than cell counting. Thirdly, in addition to the quantitative data collection, the method could be applied for identifying responsive cells. This might be very useful in identification of e.g. orexin-responding neurones in a large population of non-responsive cells in primary cultures. [source] Photopatch testing , a retrospective review using the 1 day and 2 day irradiation protocolsCONTACT DERMATITIS, Issue 2 2006R. J. Batchelor Photopatch testing is indicated in the investigation of patients with eczematous eruptions, affecting mainly light-exposed sites and in those who give a history of worsening of their condition with sun exposure. 3 different protocols are described by the British Photodermatology Group (Br J Dermatol 1997:136:371,376), 1 of which includes irradiation of allergens 1 day after application and 2 using irradiation of allergens 2 days after application. There is no evidence for superiority of any of these protocols. We reviewed the records of all patients who underwent photopatch testing in Leeds over a 50-month period, who had had the allergens applied in triplicate with 1 set irradiated after 1 day occlusion and another after 2 days. The control was occluded for 2 days. Readings were performed at 2 days and 4 days. 15 of 74 patients photopatch tested during this period had 49 positive results between them. 43 of these were felt to be of current relevance to their clinical problem. 34 of the positive results were indicative of photoallergy. Additional photoallergic reactions were detected following 2 days occlusion and irradiation compared with 1 day occlusion (16 versus 5). In conclusion, our case series suggests that 2 days occlusion before irradiation of allergens is more sensitive at detecting photoallergy. [source] DNA extraction procedure: a critical issue for bacterial diversity assessment in marine sedimentsENVIRONMENTAL MICROBIOLOGY, Issue 2 2006Gian Marco Luna Summary In order to evaluate whether different DNA extraction procedures can affect estimates of benthic bacterial diversity, based on 16S rRNA gene terminal restriction fragment length polymorphism (T-RFLP) fingerprinting technique, we compared two in situ lysis procedures (a SDS-based protocol and a commercial kit for DNA recovery) and one cell-extraction protocol on a variety of marine sediments. Despite the two in situ lysis procedures resulted in significantly different DNA yields (highest with the SDS in situ lysis), estimates of bacterial diversity provided a not significantly different ribotype richness, as well as similar values of the Shannon-Wiener (H,) and Margalef (d) indices of biodiversity and of evenness (Pielou index, J). Conversely, the cell-extraction procedure for DNA extraction resulted always in a significantly lower ribotype richness and diversity. The analysis of similarities (anosim) among the T-RFLP electropherograms allowed concluding that ribotypes composition did not change significantly using different protocols. However, the analysis of ,-diversity (turnover diversity) revealed that a large number of ribotypes was observed exclusively with one of the three protocols utilized. When unshared ribotypes from in situ lysis and cell extraction were pooled together, total ribotype richness resulted much higher (up to 80%). Our results indicate that estimates of ribotype diversity based on a single protocol of DNA extraction can significantly underestimate the total number of bacterial ribotypes present in the benthic domain. We recommend that future studies will not only integrate different DNA extraction procedures, but also will explore the possibility of integrating two or more different genetic markers in order to increase our ability to detect the actual bacterial diversity in environmental samples. [source] Differences between the effects of three plasticity inducing protocols on the organization of the human motor cortexEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 3 2006Karin Rosenkranz Abstract Several experimental protocols induce lasting changes in the excitability of motor cortex. Some involve direct cortical stimulation, others activate the somatosensory system and some combine motor and sensory stimulation. The effects usually are measured as changes in amplitude of the motor-evoked-potential (MEP) or short-interval intracortical inhibition (SICI) elicited by a single or paired pulses of transcranial magnetic stimulation (TMS). Recent work has also tested sensorimotor organization within the motor cortex by recording MEPs and SICI during short periods of vibration applied to single intrinsic hand muscles. Here sensorimotor organization is focal: MEPs increase and SICI decreases in the vibrated muscle, whilst the opposite occurs in neighbouring muscles. In six volunteers we compared the after effects of three protocols that lead to lasting changes in cortical excitability: (i) paired associative stimulation (PAS) between a TMS pulse and an electrical stimulus to the median nerve; (ii) motor practice of rapid thumb abduction; and (iii) sensory input produced by semicontinuous muscle vibration, on MEPs and SICI at rest and on the sensorimotor organization. PAS increased MEP amplitudes, whereas vibration changed sensorimotor organization. Motor practice had a dual effect and increased MEPs as well as affecting sensorimotor organization. The implication is that different protocols target different sets of cortical circuits. We speculate that protocols that involve repeated activation of motor cortical output lead to lasting changes in efficacy of synaptic connections in output circuits, whereas protocols that emphasize sensory inputs affect the strength of sensory inputs to motor circuits. [source] Cerebellar contribution to spatial event processing: do spatial procedures contribute to formation of spatial declarative knowledge?EUROPEAN JOURNAL OF NEUROSCIENCE, Issue 9 2003L. Mandolesi Abstract Spatial knowledge of an environment involves two distinct competencies: declarative spatial knowledge, linked to where environmental cues are and where the subject is with respect to the cues, and, at the same time, procedural spatial knowledge, linked to how to move into the environment. It has been previously demonstrated that hemicerebellectomized (HCbed) rats are impaired in developing efficient exploration strategies, but not in building spatial maps or in utilizing localizing cues. The aim of the present study was to analyse the relationships between spatial procedural and declarative knowledge by using the open field test. HCbed rats have been tested in two different protocols of the open field task. The results indicate that HCbed animals succeeded in moving inside the arena, in contacting the objects and in habituating to the new environment. However, HCbed animals did not react to environmental changes, when their impaired explorative pattern was inappropriate to the environment, suggesting that they were not able to represent a new environment because they were not able to explore it appropriately. Nevertheless, when their altered procedures were favoured by object arrangement, they detected environmental changes as efficiently as did normal rats. This finding suggests that no declarative spatial learning is possible without appropriate procedural spatial learning. [source] Immune tolerance induction in patients with haemophilia A with inhibitors: a systematic reviewHAEMOPHILIA, Issue 4 2003J. Wight Summary., In some patients with haemophilia A, therapeutically administered factor VIII (FVIII) comes to stimulate the production of antibodies (inhibitors) which react with FVIII to render it ineffective. As a result, FVIII cannot be used prophylactically and patients become liable to recurrent bleeds. There are two elements to the management of patients with inhibitors: the treatment of bleeding episodes, and attempts to abolish inhibitor production through the induction of immune tolerance. This paper reports a systematic review of the best available evidence of clinical effectiveness in relation to immune tolerance induction (ITI) in patients with haemophilia A with inhibitors. Owing to the lack of randomized controlled trials on this topic, broad inclusion criteria with regard to study design were applied in order to assess the best available evidence for each intervention. As a result of the clinical and methodological heterogeneity of the evidence, it was not appropriate to pool data across studies; instead, data were synthesized using tabulation and qualitative narrative assessment. The International Registry provides the most reliable estimate of the proportion of successful cases of ITI [48.7%, 95% confidence interval (CI) 42.6,52.7%]. The duration of effect is unclear, but relapses appear to be infrequent. The International Registry shows a rate of relapse of 15% at 15 years. The comparative effectiveness of different protocols is uncertain, as no trials have been undertaken which compare them directly. However, the evidence suggests that the Bonn protocol may be more effective than the Malmö or low-dose protocols. There is no good evidence that immunosuppressive drug regimens are effective. [source] Lactobacillus plantarum 299V in the treatment and prevention of spontaneous colitis in interleukin-10-deficient miceINFLAMMATORY BOWEL DISEASES, Issue 2 2002Michael Schultz Abstract Interleukin (IL)-10-deficient (IL-10,/,) mice develop colitis under specific pathogen-free (SPF) conditions and remain disease free if kept sterile (germ free [GF]). We used four different protocols that varied the time-points of oral administration of Lactobacillus plantarum 299v (L. plantarum) relative to colonization with SPF bacteria to determine whether L. plantarum could prevent and treat colitis induced by SPF bacteria in IL-10,/, mice and evaluated the effect of this probiotic organism on mucosal immune activation. Assessment of colitis included blinded histologic scores, measurements of secreted colonic immunoglobulin isotypes, IL-12 (p40 subunit), and interferon (IFN)-, production by anti-CD3-stimulated mesenteric lymph node cells. Treating SPF IL-10,/, mice with L. plantarum attenuated previously established colonic inflammation as manifested by decreased mucosal IL-12, IFN-,, and immunoglobulin G2a levels. Colonizing GF animals with L. plantarum and SPF flora simultaneously had no protective effects. Gnotobiotic IL-10,/, mice monoassociated with L. plantarum exhibited mild immune system activation but no colitis. Pretreatment of GF mice by colonization with L. plantarum, then exposure to SPF flora and continued probiotic therapy significantly decreased histologic colitis scores. These results demonstrate that L. plantarum can attenuate immune-mediated colitis and suggest a potential therapeutic role for this agent in clinical inflammatory bowel diseases. [source] A retrospective comparison of outcome of root canal treatment using two different protocolsINTERNATIONAL ENDODONTIC JOURNAL, Issue 3 2000S. E. Hoskinson [source] A model of sequential effects in common pool resource dilemmasJOURNAL OF BEHAVIORAL DECISION MAKING, Issue 1 2002David V. Budescu Abstract Experimental games are often used as models of social dilemmas in which small groups of players have unrestricted access to, and share, a common and finite resource of desirable goods in the absence of any binding coordination mechanism. Examples are fishing, hunting, and use of computational facilities. Experimental studies of social dilemmas employ different protocols of play that differ from each other in terms of the information available to the players when they register their requests from the common resource. In this study we focus on the sequential protocol, where each participant has complete information about his or her position and the total requests of the previous movers, and the positional protocol, where each player only knows his or her position, but has no information about the other's requests. Previous research has found a robust position effect: individual requests are inversely related to the players' positions in the sequence with the first mover requesting most, and the last mover requesting the least. In an attempt to characterize the nature and intensity of the position effect, we developed and tested a descriptive model with one free (individual specific) parameter. The parameter is estimated from the players' requests under the positional protocol (i.e. in the absence of any information about the other's requests) and, as such, quantifies the social norm of ,advantage of the early mover'. In a range of cases examined, including different group sizes, resource pool sizes, and positions, the model predicted very accurately individuals' requests in the sequential protocol (with full information about position and others' requests). Copyright © 2002 John Wiley & Sons, Ltd. [source] Studies on the cellular uptake of substance P and lysine-rich, KLA-derived model peptides,JOURNAL OF MOLECULAR RECOGNITION, Issue 1 2005Johannes Oehlke Abstract In the last decade many peptides have been shown to be internalized into various cell types by different, poorly characterized mechanisms. This review focuses on uptake studies with substance P (SP) aimed at unravelling the mechanism of peptide-induced mast cell degranulation, and on the characterization of the cellular uptake of designed KLA-derived model peptides. Studies on structure,activity relationships and receptor autoradiography failed to detect specific peptide receptors for the undecapeptide SP on mast cells. In view of these findings, a direct interaction of cationic peptides with heterotrimeric G proteins without the participation of a receptor has been proposed. Such a process would require insertion into and translocation of peptides across the plasma membrane. In order to clarify whether a transport of cationic peptides into rat peritoneal mast cells is possible, transport studies were performed by confocal laser scanning microscopy (CLSM) using fluorescence-labeled Arg3,Orn7 -SP and its D -amino acid analog, all- D -Arg3,Orn7 -SP, as well as by electron microscopic autoradiography using 3H-labelled SP and 125I-labelled all- D -SP. The results obtained by CLSM directly showed translocation of SP peptides into pertussis toxin-treated cells. Kinetic experiments indicated that the translocation process was rapid, occurring within a few seconds. Mast cell degranulation induced by analog of magainin 2 amide, neuropeptide Y and the model peptide acetyl-KLALKLALKALKAALKLA-amide was also found to be very fast, pointing to an extensive translocation of the peptides. In order to learn more about structural requirements for the cellular uptake of peptides, the translocation behavior of a set of systematically modified KLA-based model peptides has been studied in detail. By two different protocols for determining the amount of internalized peptide, evidence was found that the structure of the peptides only marginally affects their uptake, whereas the efflux of cationic, amphipathic peptides is strikingly diminished, thus allowing their enrichment within the cells. Although the mechanism of cellular uptake, consisting of energy-dependent and -independent contributions, is not well understood, KLA-derived peptides have been shown to deliver various cargos (PNAs, peptides) into cells. The results obtained with SP- and KLA-derived peptides are discussed in the context of the current literature. Copyright © 2004 John Wiley & Sons, Ltd. [source] Effect of the preliminary hydration on gastric emptying time for water in healthy volunteersACTA ANAESTHESIOLOGICA SCANDINAVICA, Issue 2 2009T. UMENAI Background: International guidelines allow healthy patients to drink clear liquid up to 2 h before general anesthesia. Recently, MRI measurements have been used for tracking gastric volume in humans. Hence, we used MRI to examine the gastric emptying rate of water in healthy volunteers with or without prior water loading. Methods: Fifteen healthy volunteers were enrolled. The participants had MRI examinations on separate days under two different protocols: The preliminary hydration protocol (group H) and the water restriction protocol (group R). After the intake of water, MR imaging was performed every 10 min for 60 min. The gastric content was outlined as area of interest (AOI), and the AOI area of each slice was summated to calculate the volume of gastric contents. Results: The 50% reduction time of gastric volume in group R and group H was 18±9 and 16±8 min (mean±SD), respectively. There were no significant changes between the two groups. Conclusion: The gastric emptying time for water evaluated with MRI was not affected by preliminary hydration, which shows the safety of repeated oral hydrations in the pre-operative period. [source] Influence of Artificial Accelerated Aging on Dimensional Stability of Acrylic Resins Submitted to Different Storage ProtocolsJOURNAL OF PROSTHODONTICS, Issue 6 2010Lucas da Fonseca Roberti Garcia DDS Abstract Purpose: The aim of this study was to evaluate the influence of artificial accelerated aging on dimensional stability of two types of acrylic resins (thermally and chemically activated) submitted to different protocols of storage. Materials and Methods: One hundred specimens were made using a Teflon matrix (1.5 cm × 0.5 mm) with four imprint marks, following the lost-wax casting method. The specimens were divided into ten groups, according to the type of acrylic resin, aging procedure, and storage protocol (30 days). GI: acrylic resins thermally activated, aging, storage in artificial saliva for 16 hours, distilled water for 8 hours; GII: thermal, aging, artificial saliva for 16 hours, dry for 8 hours; GIII: thermal, no aging, artificial saliva for 16 hours, distilled water for 8 hours, GIV: thermal, no aging, artificial saliva for 16 hours, dry for 8 hours; GV: acrylic resins chemically activated, aging, artificial saliva for 16 hours, distilled water for 8 hours; GVI: chemical, aging, artificial saliva for 16 hours, dry for 8 hours; GVII: chemical, no aging, artificial saliva for 16 hours, distilled water for 8 hours; GVIII: chemical, no aging, artificial saliva for 16 hours, dry for 8 hours GIX: thermal, dry for 24 hours; and GX: chemical, dry for 24 hours. All specimens were photographed before and after treatment, and the images were evaluated by software (UTHSCSA , Image Tool) that made distance measurements between the marks in the specimens (mm), calculating the dimensional stability. Data were submitted to statistical analysis (two-way ANOVA, Tukey test, p= 0.05). Results: Statistical analysis showed that the specimens submitted to storage in water presented the largest distance between both axes (major and minor), statistically different (p < 0.05) from control groups. Conclusions: All acrylic resins presented dimensional changes, and the artificial accelerated aging and storage period influenced these alterations. [source] Cortisol response to two different doses of intravenous synthetic ACTH (tetracosactrin) in overweight catsJOURNAL OF SMALL ANIMAL PRACTICE, Issue 12 2000J. P. Schoeman Fifteen middle-aged to older, overweight cats attending a first-opinion clinic were investigated to rule out hyperadrenocorticism as a cause of their weight problem, using two different protocols for the adrenocortlcotropic hormone (ACTH) stimulation test. The cats received intravenous synthetic ACTH (tetracosactrin) at an initial dose of 125 ,g; a second test was performed between two and three weeks later, using a dose of 250 vg intravenously. The mean basal serum cortisol concentration was 203 nmol/litre (range 81 to 354 nmol/litre). The highest mean serum cortisol concentration occurred at 60 minutes following the 125 ,g dose and at 120 minutes following the 250 ,g dose. There was, however, no statistically significant difference between these peak cortisol concentrations attained using either dose of tetracosactrin. A significantly higher mean serum cortisol concentration was attained after the higher dose at the 180 minutes time point, indicating a more prolonged response when compared with the lower dose. The cats were followed up for one year after the initial investigations and none were found to develop hyperadrenocorticism during this time. [source] HCV-RNA In Sural Nerve From Hcv Infected Patients With Peripheral NeuropathyJOURNAL OF THE PERIPHERAL NERVOUS SYSTEM, Issue 1 2001L De Martino Objective: Evaluation of hepatitis C virus (HCV) by reverse transcription-polymerase chain reaction (RT-PCR) in peripheral nerve tissues from HCV infected patients with peripheral neuropathy. METHODS: RT-PCR was performed on homogenates of nerve biopsies from 17 consecutive HCV-positive patients with peripheral neuropathy, with or without mixed cryoglobulinemia, hospitalised from 1996 to 2000. Sural nerve specimens were frozen in iso-pentane pre-cooled in liquid nitrogen and stored at ,80°C until use. RNA was extracted from ten 7-,m thick cryostatic sections or from a nerve trunk specimen of about 3 mm length, collected from each biopsy. Three different protocols of RNA extraction were tested (1,3). Complementary DNAs (cDNAs) were obtained without or with RNasin (Promega, Madison, WI) addition in the reaction mixture to inhibit residual RNase activity. Two sets of commercially available PCR primers for the outer and the nested reaction were used. PCR products were analysed by agarose gel electrophoresis and ethidium bromide staining. Serum samples and liver specimens from proven HCV positive patients served as positive controls, whereas sera from healthy subjects were negative controls. RESULTS: Sufficient amount of RNA could be obtained either by cryostatic sections or by in toto nerve specimens. Extraction by Trizol (Gibco-BRL) allowed the best concentration and purity of RNA as assessed by biophotometry. The presence of RNasin didn't improve the cDNA synthesis. The resulting amplification product of the nested PCR was 187 bp long. We have always observed this product in our positive controls and never in the negative. Six samples from patients either with or without cryoglobulinemia resulted positive; 7 were negative. Four samples gave variable results. CONCLUSIONS: While 40% of the nerves in our series were undoubtedly HCV positive, the cause(s) of negative and variable results in the remaining samples is likely more complex than variations in the detection protocols and deserve further investigations. REFERENCES: 1) Chomczynski P, Sacchi N (1987). Anal Biochem 162:156. 2) Marquardt O et al. (1996). Med Microbiol Lett 5:55. 3) Chomczynski P (1993). Bio/Techniques 15:532. [source] Laser phototherapy as topical prophylaxis against head and neck cancer radiotherapy-induced oral mucositis: Comparison between low and high/low power lasersLASERS IN SURGERY AND MEDICINE, Issue 4 2009Alyne Simões PhD Abstract Background and Objective Oral mucositis is a dose-limiting and painful side effect of radiotherapy (RT) and/or chemotherapy in cancer patients. The purpose of the present study was to analyze the effect of different protocols of laser phototherapy (LPT) on the grade of mucositis and degree of pain in patients under RT. Patients and Methods Thirty-nine patients were divided into three groups: G1, where the irradiations were done three times a week using low power laser; G2, where combined high and low power lasers were used three time a week; and G3, where patients received low power laser irradiation once a week. The low power LPT was done using an InGaAlP laser (660 nm/40 mW/6 J,cm,2/0.24 J per point). In the combined protocol, the high power LPT was done using a GaAlAs laser (808 nm, 1 W/cm2). Oral mucositis was assessed at each LPT session in accordance to the oral-mucositis scale of the National Institute of the Cancer,Common Toxicity criteria (NIC-CTC). The patient self-assessed pain was measured by means of the visual analogue scale. Results All protocols of LPT led to the maintenance of oral mucositis scores in the same levels until the last RT session. Moreover, LPT three times a week also maintained the pain levels. However, the patients submitted to the once a week LPT had significant pain increase; and the association of low/high LPT led to increased healing time. Conclusions These findings are desired when dealing with oncologic patients under RT avoiding unplanned radiation treatment breaks and additional hospital costs. Lasers Surg. Med. 41:264,270, 2009. © 2009 Wiley-Liss, Inc. [source] Novel renoprotective actions of erythropoietin: New uses for an old hormone (Review Article)NEPHROLOGY, Issue 4 2006DAVID W JOHNSON SUMMARY: Erythropoietin (EPO) has been used widely for the treatment of anaemia associated with chronic kidney disease and cancer chemotherapy for nearly 20 years. More recently, EPO has been found to interact with its receptor (EPO-R) expressed in a large variety of non-haematopoietic tissues to induce a range of cytoprotective cellular responses, including mitogenesis, angiogenesis, inhibition of apoptosis and promotion of vascular repair through mobilization of endothelial progenitor cells from the bone marrow. Administration of EPO or its analogue, darbepoetin, promotes impressive renoprotection in experimental ischaemic and toxic acute renal failure, as evidenced by suppressed tubular epithelial apoptosis, enhanced tubular epithelial proliferation and hastened functional recovery. This effect is still apparent when administration is delayed up to 6 h after the onset of injury and can be dissociated from its haematological effects. Based on these highly encouraging results, at least one large randomized controlled trial of EPO therapy in ischaemic acute renal failure is currently underway. Preliminary experimental and clinical evidence also indicates that EPO may be renoprotective in chronic kidney disease. The purpose of the present article is to review the renoprotective benefits of different protocols of EPO therapy in the settings of acute and chronic kidney failure and the potential mechanisms underpinning these renoprotective actions. Gaining further insight into the pleiotropic actions of EPO will hopefully eventuate in much-needed, novel therapeutic strategies for patients with kidney disease. [source] Alignment of protein sequences by their profilesPROTEIN SCIENCE, Issue 4 2004Marc A. Marti-Renom Abstract The accuracy of an alignment between two protein sequences can be improved by including other detectably related sequences in the comparison. We optimize and benchmark such an approach that relies on aligning two multiple sequence alignments, each one including one of the two protein sequences. Thirteen different protocols for creating and comparing profiles corresponding to the multiple sequence alignments are implemented in the SALIGN command of MODELLER. A test set of 200 pairwise, structure-based alignments with sequence identities below 40% is used to benchmark the 13 protocols as well as a number of previously described sequence alignment methods, including heuristic pairwise sequence alignment by BLAST, pairwise sequence alignment by global dynamic programming with an affine gap penalty function by the ALIGN command of MODELLER, sequence-profile alignment by PSI-BLAST, Hidden Markov Model methods implemented in SAM and LOBSTER, pairwise sequence alignment relying on predicted local structure by SEA, and multiple sequence alignment by CLUSTALW and COMPASS. The alignment accuracies of the best new protocols were significantly better than those of the other tested methods. For example, the fraction of the correctly aligned residues relative to the structure-based alignment by the best protocol is 56%, which can be compared with the accuracies of 26%, 42%, 43%, 48%, 50%, 49%, 43%, and 43% for the other methods, respectively. The new method is currently applied to large-scale comparative protein structure modeling of all known sequences. [source] Metabolic evaluation of cooled equine spermatozoaANDROLOGIA, Issue 2 2010A. B. Vasconcelos Summary Microscopy has been used in the routine evaluation of sperm metabolism. Nevertheless, it has limited capacity to preview male fertility. As calorimetry may be used to evaluate directly the metabolic activity of a biological system, the aim of this study was to use microcalorimetry as an additive method for sperm metabolism evaluation of cooled equine semen. Two ejaculates of four stallions were collected and motility, viability (eosin 3%) and membrane functional integrity (hyposmotic swelling test) of spermatozoa were evaluated. Sperm samples were processed following different protocols and the metabolism of these samples was accessed by calorimetry. Centrifugation is part of some of these processing protocols and although this procedure has been deleterious for sperm viability and plasma membrane integrity, no decrease in sperm motility was observed. Microcalorimetry was capable of detecting the positive effect of re-suspending the sperm pellet with Kenney extender. Thus, the use of microcalorimetry offered additional information for equine sperm metabolism evaluation and was efficient in detecting important information from sperm cell metabolism. [source] Ageing in granular aluminium insulating thin filmsANNALEN DER PHYSIK, Issue 12 2009J. Delahaye Abstract We present a new set of electrical field effect measurements on granular aluminium insulating thin films. We have explored how the conductance relaxations induced by gate voltage changes depend on the age of the system, namely the time elapsed since its quench at low temperature. A clear age dependence of the relaxations is seen, qualitatively similar to ageing effects seen in other well studied glassy systems such as spin glasses or polymers. We explain how our results differ from the previous ones obtained with different protocols in indium oxide and granular aluminium thin films. Our experimental findings bring new information on the dynamics of the system and put new constraints on the theoretical models that may explain slow conductance relaxations in disordered insulators. [source] EFFECTS OF DIFFERENT SAMPLE PREPARATION METHODS ON STABLE CARBON AND OXYGEN ISOTOPE VALUES OF BONE APATITE: A COMPARISON OF TWO TREATMENT PROTOCOLS*ARCHAEOMETRY, Issue 1 2010C. J. YODER Researchers have long debated the appropriateness of stable isotope analysis of bone apatite to reconstruct the diets of ancient animals. The debate has centred, in part, on diagenesis of bone mineral from interaction with the burial environment. A number of acetic acid treatments are used to remove diagenetic carbonates from samples; however, less is known on how different protocols alter stable isotope values. We compare two common acetic acid solution treatments (0.1 M versus 1.0 M-buffered) to examine the effects on carbon and oxygen isotope values and Fourier transform infrared spectroscopy (FTIR) spectra in human bone from different burial contexts. Results indicate that both treatments have a similar effect on isotope values and FTIR spectra in bone apatite. [source] The combination of molecular dynamics with crystallography for elucidating protein,ligand interactions: a case study involving peanut lectin complexes with T-antigen and lactoseACTA CRYSTALLOGRAPHICA SECTION D, Issue 11 2001J. V. Pratap Peanut lectin binds T-antigen [Gal,(1,3)GalNAc] with an order of magnitude higher affinity than it binds the disaccharide lactose. The crystal structures of the two complexes indicate that the higher affinity for T-antigen is generated by two water bridges involving the acetamido group. Fresh calorimetric measurements on the two complexes have been carried out in the temperature range 280,313,K. Four sets of nanosecond molecular-dynamics (MD) simulations, two at 293,K and the other two at 313,K, were performed on each of the two complexes. At each temperature, two somewhat different protocols were used to hydrate the complex in the two runs. Two MD runs under slightly different conditions for each complex served to assess the reliability of the approach for exploring protein,ligand interactions. Enthalpies based on static calculations and on MD simulations favour complexation involving T-antigen. The simulations also brought to light ensembles of direct and water-mediated protein,sugar interactions in both the cases. These ensembles provide a qualitative explanation for the temperature dependence of the thermodynamic parameters of peanut lectin,T-antigen interaction and for the results of one of the two mutational studies on the lectin. They also support the earlier conclusion that the increased affinity of peanut lectin for T-antigen compared with that for lactose is primarily caused by additional water bridges involving the acetamido group. The calculations provide a rationale for the observed sugar-binding affinity of one of the two available mutants. Detailed examination of the calculations point to the need for exercising caution in interpreting results of MD simulations: while long simulations are not possible owing to computational reasons, it is desirable to carry out several short simulations with somewhat different initial conditions. [source] Blending telephony and IPTV: Building the TV-link service package using the Alcatel-Lucent Service BrokerÔBELL LABS TECHNICAL JOURNAL, Issue 1 2007Andre Beck This paper discusses the design and implementation of blended services, composite services whose base component services interact within a common control structure. The paper focuses on blended services that combine aspects of Internet Protocol television (IPTV) and IP Multimedia Subsystem, (IMS)-based telephony services and, in particular, focuses on a service that blends TV viewing and telephone call-handling functions. If a subscriber to this service receives a telephone call while watching TV, the caller ID is displayed on the subscriber's TV screen and the subscriber can signal call control instructions using the set-top box remote control. In this service blend, functions from each base service are coordinated with actions in the other. The mechanism allowing this service is an information channel that allows status and control information to move between the subscriber's set-top box and the IMS system. The services described in this paper are built upon a software foundation called the Alcatel-Lucent Service BrokerÔ platform, a platform that has unique capabilities to support such an information channel and to serve as the gateway between IMS and digital TV. This foundation offers service developers a set of important functions for the creation of blended services. Specifically, the Alcatel-Lucent Service Broker offers a means of controlling the invocation of base services, a means of communicating through different protocols, and support for large-scale deployment and for service customization. © 2007 Alcatel-Lucent. [source] | |||||||||||||||||||