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Maps & Routes
Charter of Commitment for Sustainable Mobility in the Algarve
Understanding of departure
"Sustainable Mobility" is a multidimensional concept that depends on the effort of all and aims to respond to the needs of the population (residents, visitors and tourists) to access the various activities that constitute the basic support of quality of life (housing, employment school, culture, health, sport, purchase of essential goods, etc.).
Responses take into account the most vulnerable groups in the system (low income, blind, elderly, etc.) in a commitment to universal accessibility, and the needs of efficient freight transport and urban and intermunicipal logistics.
The objective is to enable current and future generations to minimize social, environmental and financial costs, to make more efficient use of existing resources, to promote new transport technologies and new mobility services and to use renewable energy whenever possible , enabling an integrated multimodal transport system for the region and financially sustainable management models in a commitment to all stakeholders.
The municipalities and other key stakeholders in the Algarve transport and mobility system form a collaborative platform for action and endorse the "Charter of Commitment to Sustainable Mobility in the Algarve", agreeing:
in a continuous manner to achieve sustainable mobility.
To assist
in the process of transforming behaviors for greater efficiency in the use of resources in mobility / accessibility.
the various transportation modes and services with the main poles of urban and inter-city movements.
experiences for the resolution of transport and mobility problems and the implementation of measures.
innovative mobility and accessibility solutions.
continuously what progress has been made in the execution of the plan and the various actions.
the transition to a low-carbon economy, by adhering to the principles of equity and social justice.
for innovation in planning practices, aiming at optimizing existing resources to achieve cost-effective actions.
mobility / accessibility solutions, through the feasibility of new practices and / or tools / management models.
Dialogue and promote the participation
of all stakeholders and the population, aiming at concertation and social inclusion.
Where do we go?
Future Stop
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The M2 channel controversy rides again
Advanced, Allostery, Basic, Drug Resistance, Medicine, Structural Biology 3 Responses »
Most people never learn about an actual scientific controversy. Almost every "controversy" that bubbles into the public eye is manufactured, often reflecting social or ethical differences rather than genuine disagreements between experts about how different models fit to reality. Actual scientific controversies tend to be highly technical, and often concern points that lay people find to be esoteric. That doesn't mean that the issues involved aren't important, or that they're even difficult to understand. One controversy that has unfolded over the past few years and now may be over relates to a seemingly simple question. Where do adamantane drugs bind to the influenza A M2 channel?
Previously, on As the Channel Twists…
Bill DeGrado and James Chou, whose
competing structures began the controversy
The M2 proton channel plays an essential role in the life cycle of the influenza virus. The activity of the channel could be blocked, at least in influenza A, by drugs called adamantanes, including amantadine and rimantadine. Unfortunately, these antiviral drugs have been fading in efficacy due to the spread of an S31N mutation that interferes with their binding. On January 31, 2008, two articles appeared in the scientific journal Nature showing adamantanes bound to the M2 channel. Unfortunately, the structures had different answers about where the drug was bound. The X-ray crystal structure from Bill DeGrado's group at the University of Pennsylvania placed amantadine in the center of the channel's pore, suggesting a simple pore-blocking model (PBM) for inhibition. The NMR structure from James Chou's group at Harvard University located rimantadine on the outside of the channel, ultimately giving rise to an allosteric, dynamic quenching model (DQM) of adamantane activity.
As outlined in my previous post on the M2 channel, there was conflicting functional evidence as to which site was actually relevant in vivo, and reasons to doubt the conclusions from both structures. Since that time, several papers have been published that substantially clarify the issue. At this point, the evidence strongly supports the PBM as an explanation of adamantane activity in vivo.
Sure adamantanes bind there, but does it matter?
The direct observation of NOEs, even weak ones, between the adamantane and the protein proved that the drugs were binding at the DQM site, but there were some significant areas of concern with this finding. The greatest worry was due to the extremely high concentration of ligand used in the NMR experiment. This opened up the possibility that the DQM site was a low-affinity site that would not see binding under normal circumstances. Because both models had explanations for the efficacy of the S31N mutation, the only way to address the question would be to make mutations that would abolish binding at the DQM site and see if adamantanes were still effective. Because aspartate 44 was proposed to form a hydrogen bond to rimantadine, it was thought that a D44A mutation would eliminate binding, and if DQM was true, adamantane activity. This prediction was borne out by an experiment performed in liposomes by the Chou lab (6), but Robert Lamb's group from Northwestern University was not able to replicate this result in X. laevis oocytes (4).
Robert Lamb has studied the
M2 channel since the 80s.
What Lamb's group did do was test different parts of the influenza A channel for adamantane sensitivity by fusing them to the adamantane-insensitive influenza B channel. These A/B M2 chimeras should in principle have adamantane susceptibility if the legitimate binding site got imported from A to B. Their first results in this experiment were somewhat inconclusive. Adding the N-terminal portion of the A channel to the C-terminal portion of the B channel (essentially sticking the PBM site into B M2) created a chimera that was somewhat sensitive to amantadine treatment, but the effect was nowhere near what occurred for WT A channel (4). Subsequently, Lamb's group expanded these experiments to add a little bit more of the N-terminal sequence to the chimera, which then almost perfectly matched the WT A channel's susceptibility. Notably, when they made the opposite chimera that incorporated the DQM site from A into the B channel, only a very slight inhibitory activity was observed upon addition of amantadine (5). While the conclusions that can be drawn from the chimeras are limited by their particularly odd provenance, the fact that transplanting the PBM site from one channel to another confers adamantane susceptibility suggests that this is the functional binding site.
An upcoming paper in PNAS clarifies the picture somewhat using surface plasmon resonance (SPR) (7). This technique detects the binding of a ligand as a change in physical force exerted by a protein tethered to the surface of a gold chip. In this case, the tethering was mediated by a DMPC liposome. This was a tricky experiment because adamantanes like the greasy portions of lipid bilayers, so they can bind to the liposome itself. Rosenberg and Casarotto, however, were able to control for this effect. Their SPR experiments detect two distinct adamantane binding sites on M2 with vastly different affinities. Rimantadine binding at the high-affinity site could be abrogated by S31N and V27A mutations, but not a D44A mutation. At the low-affinity site, rimantadine binding could be knocked out by a D44A mutation or an S31N mutation, but not a V27A mutation. This result indicates that both binding sites are functional (and, incidentally, that the S31N mutation does indeed exert an allosteric effect on the DQM site). However, the authors note that the adamantane concentrations used in actual treatment are too low to significantly populate the low-affinity site, given the dissociation constants they calculated. This argues that the DQM site is irrelevant in vivo.
Amantadine caught in the pore
Mei Hong has studied M2
extensively by NMR
One of the problems with the PBM was that the crystal structure that supported it was unsatisfactory in a variety of ways. The structure was made using a construct that consisted of only the transmembrane segment of the protein. This construct could not be reconstituted in micelles, and functional experiments showed that it was not very similar to the WT in terms of its activity. In addition, the extra electron density in the pore could not be unambiguously assigned as amantadine. In a paper from February of this year, the DeGrado group collaborated with Mei Hong's group at Iowa State University to produce a structure of amantadine bound to M2 using solid-state NMR (2). An important advantage of this approach is that one can take spectra of proteins embedded in a membrane without penalty, because there is no requirement for the protein to tumble freely. While there are some trade-offs in terms of resolution and the kinds of data that can be obtained, biomolecular solid-state NMR can help us answer some very tricky questions.
Amantadine (yellow) in the
pore binding site.
The approach proved to be especially fruitful here. Cady et al. used a technique that allowed them to determine whether the amantadine was near a particular residue, by labeling residues with 13C and the amantadine with 2H. If a 13C nucleus is coupled to a 2H nucleus by a dipolar interaction, a pulse that dephases the 2H nucleus will affect the 13C nucleus in a distance-dependent manner. When Cady et al. made samples at a ratio of one amantadine per channel, they found that the signal from S31 was significantly broadened, but that from D44 was not, proving that the amantadine is close to S31 under these conditions. The D44 signal was affected at higher amantadine concentrations, but never to the same degree as the S31 signal. Other residues at the PBM site were also affected by the presence of amantadine. Using an alternative version of the REDOR experiment, Cady et al. were able to generate distance constraints and, in combination with other data, generate a structure for the channel with amantadine bound (see their figure at right) (2). Note the residues that are close to the amantadine in this structure: V27, S31, G34, and H37. This will be important in a minute.
The Cady et al. paper has several advantages over the DeGrado group's original crystal structure. The first, and most important, is that the protein was reconstituted in DMPC vesicles rather than OGP bilayers. The lipids themselves, and the vesicle structure, more closely mimic the likely environment in vivo than the crystal conditions, and they are also more biologically-relevant than the DHPC micelles employed in Chou's original determination. In addition, the pH of 7.5 matches the experimental conditions used by Chou and allows for a direct comparison of results under conditions where the channel's conformation should in principle be the same. The REDOR results provide unambiguous evidence that amantadine binds preferentially in the pocket for this construct. Their observation that amantadine has approximately 100-fold higher affinity for the PBM site, but can also bind the DQM site, agrees nicely with the functional data, especially the SPR results.
Support for an allosteric mechanism?
Bob Griffin,
SSNMR master
However, Cady et al. still used the truncated construct that possesses significantly altered activity relative to WT, which brings us to an upcoming paper in JACS by Andreas et al. (1). Chou's group collaborated with Robert Griffin's group at MIT to map the chemical shifts of a somewhat larger construct of M2 that is known to have relatively normal activity. The longer construct is also known to form a fairly stable tetramer, and this may have improved its spectral properties. The construct was inserted into membrane bilayers that were lyophilized for the NMR experiment, with and without rimantidine present. The chemical shifts of 15N and 13C nuclei of residues in the transmembrane helices were compared between these two states as a way of localizing the adamantane binding site. As Andreas et al. show in their figure 2 (recapitulated below in a slightly altered fashion), the chemical shift changes in response to the binding event are quite widespread. The authors argue that this observation favors an allosteric inhibition mechanism. That may well be true, in a sense, but unfortunately that does not mean that the observation favors DQM.
The binding of a ligand generally alters the chemical shifts of a protein by changing its structure, reshaping the arrangement of bond angles that dictates the electron distribution around the relevant nuclei. It is of course possible that a small change in the protein's conformation at the binding site propagates into a large change elsewhere, but this is not typically observed. The most reasonable interpretation of chemical shift changes (Δδ) upon ligand binding is that the largest shifts observed are nearest to the ligand, while the smaller shifts are farther away. The original version of figure 2 presents the data in a fairly simple way, and does not distinguish very large changes from relatively small ones. So, I made a new version of the figure for you, where I've taken the additional step of scaling the color according to the magnitude of Δδ.
M2 channel showing Δδ
due to rimantidine binding
This adaptation of the Andreas et al. figure, shows the NMR micelle structure (PDB code: 2RLF) with purple rimantadine at the DQM binding site and the helices colored in blue according to the Δδ. This was done very roughly, by eyeballing the graphs to the left in figure 2, scaling the Δδ based on the nucleus measured, and adding it all up across all three nuclei. The more intense the blue, the larger the Δδ. It should be immediately apparent that the largest chemical shift changes are located a substantial distance from the DQM site (although there are some smaller changes near that site). What's perhaps less immediately obvious is that the largest chemical shift changes belong to residues located in the pore. To make this point more clear, check out the figure below and to the left. This is the same structure, that I have now tilted so we are looking down the barrel of the channel. The side chains of the four residues with the largest chemical shift changes (V27, S31, G34, H37) are shown in light green for contrast (obviously there's nothing for G34). Three of them are sticking into the pore in this model (G34′s Cα faces the pore); the fourth is S31. You might recall that the structure from Cady et al. (above) puts all four of these large Δδ residues right next to the ligand.
The largest chemical shift changes in response to rimantidine binding occur near the proposed PBM site and far from the DQM site, and the residues most affected are those facing the pore. Andreas et al. argue that this information is insufficient to positively localize the drug, due to the large chemical shift change at H37 Cα, but this doesn't seem particularly convincing. The concentration of large chemical shift changes at the N-terminal end of the channel strongly argues for the ligand binding in this region. As for the significant Δδ at H37, the observations could quite possibly be due to ring-current effects from the repositioning of the adjacent tryptophan side chains (light red in the figure to the left). We know that binding of adamantanes has at least some effect on W41 thanks to Czabotar et al. (3), who measured adamantane binding by observing changes in intrinsic tryptophan fluorescence. Of course, changes in fluorescence are a very general indicator of structural or dynamic change, but the previous finding supports the possibility of interpreting the H37 observations as side-chain effects rather than ligand proximity.
In contrast, there is no such explanation for the significant chemical shift changes at the N-terminal end of the channel, which has no aromatic residues. The significant Δδ in this region must be due to rearrangements of these residues themselves, rather than long-range effects or ring currents. Thus, the most plausible model explaining these data remains one in which adamantanes bind near V27 and S31, propagating some kind of structural change to W41, rather than the other way around. In this regard, I agree that the data from figure 2 establish that adamantane binding has an allosteric effect. These data, however, do not support the DQM Chou proposed previously.
Conclusions, Lessons
The DQM hit its high-water mark with the Pielak et al. PNAS paper back in 2009 (6), in which the model's prediction that the D44A mutation would significantly alter adamantane sensitivity was borne out by experiments in liposomes. Since that time, however, the evidence has started to weigh mostly against it. The D44A results could not be replicated in X. laevis oocytes, and lovely chimera experiments in this system demonstrated that the N-terminal region of M2 was critical for adamantane sensitivity (4)(5). Live viruses remained sensitive to adamantanes even if they were reverse-engineered to have the D44A mutation. Rosenberg and Casarotto showed that the D44A mutation only affected binding at absurdly high rimantadine concentrations (7). Finally, the Cady et al. study provided unambiguous evidence for adamantane in the pore of the channel (2). In light of these findings, only a crystal-clear result in favor of the DQM could really save it.
Although their findings convincingly illustrate an allosteric effect from rimantidine binding, Andreas et al. do not provide that result. Even their own chemical shift data seem to support the PBM model. Of course direct dipolar couplings would provide a totally unambiguous answer as to the location of the rimantidine, but in light of our existing knowledge about the system, that experiment doesn't seem necessary. At this point there is no serious reason to doubt that the physiological inhibition of M2 channel results from adamantane drugs binding to the pore.
The papers cited in this article represent decades of man-hours and significant amounts of money spent in resolving what might seem like an esoteric point. Given the enormous effort that went into resolving the seemingly simple question, you might be tempted to ask what went wrong. The answer is, "nothing". This is how the scientific process is supposed to work. Two groups came at the same problem in different ways and got different answers, which is hardly a surprise because no experiment is perfect. More experiments were carried out to determine which model best represented the physical reality. Eventually, the weight of the evidence strongly supported one model over the other. The best data we have right now really point to a single conclusion. The process succeeded, and nobody needed a superior court judge or a congressional hearing.
That doesn't mean we can't take some lessons from the experience. Most prominent among these is that we must have serious reservations about NMR structures derived from proteins bound to or inserted in micelles. What we know about the M2 channel tells us that adamantanes prefer to bind in the pore. That they did not do so (or at least, could not be detected doing so) in the micelle-based structure suggests that something about the micelle itself made that impossible. We know that the forces exerted on proteins by membrane curvature can be substantial, and the structure of a micelle is very unlike the structure of a cell membrane. Solution NMR in bicelles may yet prove to be a superior approach for some systems, but in this case it was solid-state NMR that provided the vital evidence. Solid-state has its own set of limitations, but it's clear proper membrane context is absolutely vital to getting good answers about membrane protein structure and function.
Knowing the actual binding site of adamantanes may prove to be very important in aiding the design of alternative drugs that achieve the same inhibition of the channel. The papers from the DeGrado and Hong groups have already made several interesting recommendations in this regard. Even what has been learned about the remote site may not be fruitless. Though it is not the source of the physiological activity of adamantanes, several experiments have made it clear that there is some kind of allosteric interaction between S31 and the DQM site. It may be possible to attack M2 through this site with a specifically-designed high-affinity drug, even if adamantanes themselves don't work this way. If that proves to be the case, then this will be the best kind of scientific controversy: one where we learn something important from both sides.
(1) Andreas, L., Eddy, M., Pielak, R., Chou, J., & Griffin, R. (2010). "Magic Angle Spinning NMR Investigation of Influenza A M2: Support for an Allosteric Mechanism of Inhibition." Journal of the American Chemical Society DOI: 10.1021/ja101537p
(2) Cady, S., Schmidt-Rohr, K., Wang, J., Soto, C., DeGrado, W., & Hong, M. (2010). "Structure of the amantadine binding site of influenza M2 proton channels in lipid bilayers." Nature, 463 (7281), 689-692 DOI: 10.1038/nature08722
(3) Czabotar, P., Martin, S.R., & Hay, A.J. (2004). "Studies of structural changes in the M2 proton channel of influenza A virus by tryptophan fluorescence." Virus Research, 99 (1), 57-61 DOI: 10.1016/j.virusres.2003.10.004
(4) Jing, X., Ma, C., Ohigashi, Y., Oliveira, F., Jardetzky, T., Pinto, L., & Lamb, R. (2008). "Functional studies indicate amantadine binds to the pore of the influenza A virus M2 proton-selective ion channel." Proceedings of the National Academy of Sciences, 105 (31), 10967-10972 DOI: 10.1073/pnas.0804958105 OPEN ACCESS
(5) Ohigashi, Y., Ma, C., Jing, X., Balannick, V., Pinto, L., & Lamb, R. (2009). "An amantadine-sensitive chimeric BM2 ion channel of influenza B virus has implications for the mechanism of drug inhibition." Proceedings of the National Academy of Sciences, 106 (44), 18775-18779 DOI: 10.1073/pnas.0910584106
(6) Pielak, R., Schnell, J., & Chou, J. (2009). "Mechanism of drug inhibition and drug resistance of influenza A M2 channel." Proceedings of the National Academy of Sciences, 106 (18), 7379-7384 DOI: 10.1073/pnas.0902548106
(7) Rosenberg, M., & Casarotto, M. (2010). "Coexistence of two adamantane binding sites in the influenza A M2 ion channel." Proceedings of the National Academy of Sciences DOI: 10.1073/pnas.1002051107
Posted by Sparky at 12:17 AM	Tagged with: influenza
Do metamorphic proteins mediate evolutionary structural transitions?
Basic, Evolution, Structural Biology Comments Off
On several previous occasions on this blog I've discussed proteins that undergo significant changes in structure without drastic changes in their primary sequence or solution conditions. In some cases, a few mutations can take a protein to a novel fold, as with Philip Bryan's protein G work. In others, closely related sequences within a whole family populate different kinds of folds, as Matt Cordes illustrated for the case of Cro proteins. In addition, there are some cases such as lymphotactin, where interconversion between two very different structures takes place at equilibrium, as illustrated by Brian Volkman's research. Each time stories like this come up I have mentioned that this kind of behavior (termed "metamorphism" in a 2008 commentary by Alexey Murzin) suggests a means by which proteins could evolve from one structure to another without losing foldedness or function. Recently, a group from the Weizmann Institute published results in PNAS that speak to this possibility.
Yadid et al. are looking at a class of proteins called β-propellers. Characteristics of the sequences of these proteins, especially high homology between different blades within a protein, suggest that these proteins are "built up" by gene duplication and fusion from precursors that were either multimeric in nature or made from a smaller number of blades (or both). In particular, they worked with a protein called tachylectin-2, that binds sugars. You can see its structure at right (or examine it at the PDB). The color-coding recognizes that the N and C termini are adjacent to one another, meaning that each "blade" of the propeller actually incorporates one strand from a neighbor. The whole protein is a bit under 250 residues in size. Previously, the authors of this paper had randomly chewed up the tachylectin-2 DNA from either end, a process that one might expect would produce a bunch of useless garbage. Some of the products of this experiment, however, were functional pentamers. But, they were about 100 amino acids long, suggesting that each monomer incorporated two blades. This meant that the structure of the pentamers had to differ from that of the original protein in some key way, but the proteins could not be crystallized due to low yields and instability.
To solve this problem, Yadid et al. performed directed-evolution refinement of the sequences of two promising candidates. From the pool of proteins thus produced they were able to crystallize two, which had interesting properties. As one might expect given the known data, both these proteins formed ten-bladed propellers (the structures have PDB codes 3KIF and 3KIH) in the form of two five-bladed propellers that were linked to each other. In the case of the mutant called Lib2-D2-15, 3 of the 100-residue subunits contributed two blades to the propeller apiece, while the remaining two monomers each contributed to three blades. That doesn't add up to 10 because each of the two oddballs contributed four strands to one blade, three strands to another, and one strand to the third. That means one blade was uniquely of that monomer and the other two were shared. One of these blades was shoved into the second propeller, generating an asymmetric pentamer. Note also that the two oddballs weren't equivalent: one was arranged 4-3-1 and the other 3-1-4. The other mutant, Lib1-B7-18, was even weirder in some ways. In that mutant, four of the monomers contributed to three blades in a 1-3-4 manner. The last monomer, however, contributed to four blades, two from each propeller, with a pattern of 1-3-1-3. Because these structures cannot form unless the monomers adopt multiple structures (3 in the case of Lib2-D2-15 and 2 in the case of Lib1-B7-18), it follows that the monomers must be metamorphic.
The evolved fragments didn't have higher stability to guanidinium hydrochloride than the source fragments, suggesting that the improved expression and solubility was not due to improved stability. The authors argue that the improved expression was mostly due to a change in the isoelectric point of the mutants, which decreased towards neutral in both cases. However, the evolved fragments also were able to refold from the denatured state, which the source fragments could not do. To the authors, this suggests that the directed evolution process actually selected for metamorphism; that is, the proteins were stabilized by an increased ability to sample states that formed productive pentamers.
The observations in this study, although very interesting, do not tell us anything directly about the evolution of tachylectin-2. While it is possible that the current protein evolved from a metamorphic precursor like these fragments, there is no direct evidence that this is the case, nothing to indicate that evolution performed in reverse what Yadid et al. did here. Whether metamorphism contributed to the diversity of β-propeller folds in general, or to the evolution of this protein in particular, remains very much an open question. The metamorphism observed here may be more a consequence of a particularly robust fold tending towards its original state than evidence of hidden metamorphic potential in singular structures. As such, the article's title strikes me as overbold given the data. That said, it is certainly not implausible that odd assemblies like these pentamers played a role somewhere along the evolutionary path that created the rich library of β-propellers we have today, and this study establishes that even very strange steps along the way can occur without destroying the protein's function or certain gross features of its structure.
Yadid, I., Kirshenbaum, N., Sharon, M., Dym, O., & Tawfik, D. (2010). "Metamorphic proteins mediate evolutionary transitions of structure." Proceedings of the National Academy of Sciences, 107 (16), 7287-7292 DOI: 10.1073/pnas.0912616107
Posted by Sparky at 2:39 AM
Dynamics conservation in the Ras superfamily
Advanced, Biomolecular Dynamics, Evolution, Structural Biology Comments Off
The proposition that general fold architecture is preserved within a family of evolutionarily-related proteins is not controversial. The amino acid sequence of a protein determines its structure, and countless studies have substantiated the idea that proteins with similar sequences will adopt similar folded conformations. Because structure and dynamics are intrinsically linked, one could reasonably assume that many features of a protein's dynamics get conserved along with the fold. A growing number of experiments show that this is indeed the case, including a recent paper in Structure (1).
We already have some evidence of fold-dependent dynamics. An NMR study from my mentor Andrew Lee's lab comparing fast fluctuations of side chains among three related proteins from the PDZ family suggested that motions on this timescale could be evolutionarily conserved (2). That study compared the model-free order parameters of methyl groups from one protein to those of their counterparts in other PDZ domains. Predicting an order parameter using dynamics data from a structurally equivalent residue in another protein was shown to be slightly more accurate than calculations from structural considerations such as packing or methyl type. In a similar vein, I have previously discussed studies on adenylate kinase enzymes from E. coli and a thermophilic organism that show they have similar backbone dynamics under conditions where their enzymatic activity is about equal, although they differ substantially from each other at room temperature.
Of course, these studies were limited and involved just a few proteins, because getting experimental data about dynamics is costly and time-consuming. For comparisons across large numbers of different proteins, computational approaches may therefore be of great value. Previously, other groups have made use of short molecular dynamics simulations or normal mode analysis. Raimondi et al. continue in this vein, combining normal-mode analysis of single structures with principal component analysis of a large set of structures from the Ras superfamily of proteins.
The Ras superfamily encompasses several groups of related folds with nucleotide-dependent activity. When GTP is bound to them, they are active and propagate a particular signal. Over time, the GTP gets hydrolyzed to GDP and the signal turns off. This catalytic process is pretty inefficient, but it can be enhanced by the action of a GTPase Activating Protein (GAP). The exchange of GDP for GTP can be enhanced by the action of a Guanine nucleotide Exchange Factor (GEF). The GTP/GDP state manifests primarily in the positioning of two loops, termed the switch regions (SwI and SwII). This mechanism allows for several different modes of control, so the Ras architecture has been repurposed many times throughout evolution for a variety of different roles.
Because the different members of the superfamily play key roles in their respective pathways, there are many structures available, often in several different states (GTP-bound, GDP-bound, GEF-bound, etc.). Raimondi et al. aligned these structures using the common features of the Ras fold and used PCA to identify flexibility across this evolutionary ensemble. The goal of PCA is to take a dataset with many potentially correlated data points (in this case, the relative positions of the backbone Cα atoms) and identify a small set of variables that explain as much of the variance as possible. Here, the principal components (PC) are expected to describe the structural variability of the fold.
The first PC, which is expected to explain the largest amount of the variability, can separate the structures by their families. That is, the displacement along PC1 can distinguish a Rho family domain from an Arf family domain. The authors call this variability function-independent, because this principal component doesn't seem to make any meaningful distinction between the GTP/active and GDP/inactive states. That appears to be a property of the second PC, which for some families does a very good job of separating the GTP from the GDP-bound forms (for others there appears to be more mixing). According to this analysis, function-dependent variability appears to be confined to one half of the protein, while function-independent variability seems to be distributed across the whole fold.
The authors also performed normal mode analysis on individual proteins from the Ras superfamily using an elastic network model. In this kind of simulation the protein is modeled as a group of Cα "nodes" connected by spring-like harmonic potentials representing covalent and non-covalent interactions. Although any one of these "bonds" can be stretched, compressed, and moved, such deformations exert a force on other bonds connected to the nodes involved, which tends to damp most motions. Certain collective deformations will be favored as a result, and these can be calculated as "normal modes" that probably reflect slow fluctuations of the fold.
The deformations detected by ENM for all individual proteins overlapped significantly with the second PC identified in the evolutionary analysis. That is, the conformational variability of a conserved domain over evolutionary time is correlated with the conformational fluctuations of a single domain on a biological time scale. This makes sense, especially in this case, because the switch regions are areas of significant conformational variability, and are connected with the conserved catalytic function of these proteins. The fact that PC1 doesn't line up with the low-frequency normal modes probably means that the conformational transitions between different family members cannot be mimicked by ordinary thermal motion, i.e. the fold cannot change this way without the aid of mutations.
Although the results in these studies might seem rather pedestrian and expected, I find them quite encouraging. We're not particularly good at predicting structure from sequence yet, and our understanding of protein dynamics is even more primitive. What these studies indicate is that it should be possible to predict the conformational fluctuations of a given protein or domain using our knowledge of a related, homologous protein. This could have positive consequences for fields such as rational drug design and protein design, which have met with limited success in part, perhaps, because they do not sufficiently account for a protein's structural fluctuations.
(1) Raimondi, F., Orozco, M., & Fanelli, F. (2010). Deciphering the Deformation Modes Associated with Function Retention and Specialization in Members of the Ras Superfamily. Structure, 18 (3), 402-414 DOI: 10.1016/j.str.2009.12.015
(2) Law, A., Fuentes, E., & Lee, A. (2009). Conservation of Side-Chain Dynamics Within a Protein Family. Journal of the American Chemical Society, 131 (18), 6322-6323 DOI: 10.1021/ja809915a
Posted by Sparky at 10:00 PM
A single residue dictates a fold
Anfinsen's dogma — that the amino acid sequence of a protein uniquely determines its structure — naturally leads one to the idea that identity between amino acid sequences means identity between structures. This has proven to be a successful paradigm: sequence similarity reliably predicts structural and functional similarity. Evidence accruing in recent years, however, suggests that for small proteins, at least, this assumption may not be entirely safe. Adding to this view, in an article in PNAS this week (it's open access, so open it up), a research team led by Philip Bryan reports that they are able to generate significantly different folds with divergent functions from sequences that differ by a single amino acid.
The researchers in this study took two independently folded domains from streptococcal protein G that have different structures and sequences, called GA and GB, which bind human serum albumin (HSA) and part of an antibody (IgG), respectively. As you can see from several figures in this paper, GA has an all α-helical structure, while GB has a helix and a four-strand β-sheet. In previous papers they have undertaken a process of slowly mutating these two domains towards each other, most recently generating a pair of proteins they called GA88 and GB88 (which I have mentioned before) that retained their original structures and functions despite differing in their amino acid sequence at only seven places (i.e. they were 88% identical). Beyond this point, mutating the sequences towards each other causes the protein stability to degrade. However, they manage to complete the project by generating sequences that are 91, 95, and 98% identical while retaining much of their original function.
GA98 and GB98 differ only in the identity of the amino acid at position 45, which is leucine in GA98 and tyrosine in GB98. The tyrosine seems to be critical in stabilizing a hairpin turn in the GB structure, possibly by interactions with an adjacent aromatic residue (F52 – see Fig. 6). In the structure of the GA95 protein (Fig. 5), L45 is partially solvent exposed, and therefore probably does not contribute substantially to stability. Of course, as a leucine is also hydrophobic, one might expect that it would also interact favorably with F52. This seems to be the case, as the GA98 protein binds to both HSA and IgG, suggesting that it transiently samples the GB structure.
As with studies I have talked about previously on lymphotactin, which has two structures in its native state, and the cro proteins, which can have different structures despite similar sequences, this research indicates that the relationship between sequence space and fold space may not be as straightforward as previously believed. We have known for some time that distance in sequence space does not imply distance in fold space — the convergent evolution of numerous proteins has shown us that, at least. These experiments inform us that distance in fold space need not imply distance in sequence space, that it is possible for a protein to undergo a structural saltation in which a single mutation gives rise to a dramatic difference in structure.
It remains to be seen whether this is true in larger proteins as it seems to be in small proteins and domains. If it is generally the case that proteins can drift towards new folds without losing their native function, then "jump" over to a completely different structure, without transiting the molten-globule state, then our understanding of evolution, particularly early evolution, may change significantly. For instance, the objection that proteins could not evolve new functions without wandering into the unfolded state loses its weight. Moreover, given that in two of these cases the proteins actually sample both structures (and hence, both functions), it is not even necessarily true that a protein must lose its original function before evolving a new one. Equilibrium switching between structures would allow a protein to fulfill both the old and new functions, albeit likely at reduced efficiency. As I've mentioned before, one answer to this lack of efficiency would be altering protein concentration by adjusting gene dosage, i.e. gene duplication. From that point, the generation of fold and functional diversity by evolution might be a straightforward process.
Alexander, P., He, Y., Chen, Y., Orban, J., & Bryan, P. (2009). From the Cover: A minimal sequence code for switching protein structure and function Proceedings of the National Academy of Sciences, 106 (50), 21149-21154 DOI: 10.1073/pnas.0906408106 OPEN ACCESS

redpajama_set_name: RedPajamaCommonCrawl


The holidays are a festive time that people love decorating their homes to bring cheer to family and friends. While it is fun to hang lights, tinsel, and so much more, it is important to be aware of potential dangers associated with decorating. In the event that you or someone you know has an injury, a Provo personal injury attorney should be contacted.
If your home has a fireplace, it's no doubt that it will be decorated with a variety of adornments. As time is enjoyed by the fireplace, it is important to be aware if your decorations are flammable. It only takes a small spark coming in contact with a small piece of flammable material to start a fire and cause injuries for you and your loved ones.
Another way fire can cause injuries is by use of candles. When not used properly, candles can cause burns, or worse, start fires. If you plan on using candles, be sure to have enough space for them to burn and keep them away from any free-flowing objects. Also, keep children and pets away from them and extinguish them once they will not be attended.
As lights are placed around your home, inside and outside, be mindful of the condition of the lights and that there are no frayed cords or wires. Also, unplug lights when you are away to avoid the possibility of an electrical fire.
When using a ladder, do not climb to heights that exceed the ladder's capacity and always have someone nearby that can spot you while the ladder is in use.
Even if all precautions are taken, injuries can still occur, possibly due to the malfunctioning of objects you use to decorate. A Provo personal injury lawyer can determine if someone else is at fault.
If you need a personal injury lawyer Orem, contact our office for a consultation.


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Cambrils és una de les nou entitats de població en què es divideix el municipi d'Odèn, a la comarca catalana del Solsonès.
 Tot i que no es pot parlar que tingui un nucli de poblament agrupat, sí que a l'entorn de l'antic castell de Cambrils i de l'església parroquial hi ha un grup d'edificis dispersos (majoritàriament masies) que, encara que no tinguin cap estructura urbana, permeten que se'l pugui considerar com un poble. En qualsevol cas, és en aquest nucli on hi ha l'ajuntament, cosa que fa que Cambrils pugui ser considerat com la capital del municipi.
Situat a l'entorn dels 1.135 m d'altitud, és al sud-oest del Tossal de Cambrils (1.803 m alt) i sota el veïnat de Llinars prop del qual hi ha la Font Salada en la qual neix la Ribera Salada (que en el seu primer tram també és coneguda amb el nom de Riu Fred) que deixa a la dreta l'església parroquial de Sant Martí (consagrada el 1051), passa pel molí de Cambrils i pel molí de la Sal (o salí de Cambrils), on es preparava la sal treta del riu.

Demografia

Història i etimologia del topònim
El lloc és esmentat ja el 839 en l'acta de consagració de la catedral de la Seu d'Urgell amb el topònim Kabrils, que segons el Diccionari català-valencià-balear prové del terme llatí caprīles que significa el lloc on s'arredossen les cabres. A l'època medieval, la senyoria pertangué als vescomtes de Cardona

En un document de 1065 hi apareix escrit Chabrils

En un mapa publicat a Roma l'any 1619 hi apareix escrit Cambrills

Referències

Vegeu també

 Roure de la Comella

Entitats de població d'Odèn

redpajama_set_name: RedPajamaWikipedia


text:
The Marathon Running Podcast
Episode #21 - How to fuel for a marathon with John Davis
All you ever wanted to know about marathon fueling with John Davis - author, coach, and PhD student
John Davis is a PhD student in biomechanics at Indiana University. He's also an author and coach. Today John breaks down the science behind optimal marathon fueling. We discuss questions such as: how many calories does it take to run a marathon? How many of those calories are carbs? How many carbs can you realistically absorb during a marathon? What do different fueling products have to offer, and which is best for me? Is it possible to become "fat adapted" and avoid the need for fueling during the race? What are my electrolyte and hydration needs during a marathon? John brings his depth of experience and knowledge of the scientific literature to bear on all these topics and makes the complex physiology accessible to all runners in practical terms.
Check out John's website: https://runningwritings.com
Sign up for John's newsletter: https://runningwritings.com/news
Check out John's book: https://www.amazon.com/Modern-Training-Physiology-Long-Distance-Runners/dp/0615790291
Follow John on Twitter: @JDruns
Follow the show on Instagram: @marathonpodcast


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Distinction of the Division II of Biological and Agricultural Sciences of the Polish Academy of Sciences for the team of prof. Bartoszek-Pączkowska | Faculty of Chemistry at the Gdańsk University of Technology
Distinction of the Division II of Biological and Agricultural Sciences of the Polish Academy of Sciences for the team of prof. Bartoszek-Pączkowska
In the photo from the left: Izabela Koss-Mikołajczyk, PhD, Eng., prof. Agnieszka Bartoszek-Pączkowska, PhD, DSc, Eng. Photo: J. Kwiatkowski / FCh Gdańsk Tech
Prof. Agnieszka Bartoszek-Pączkowska, PhD, DSc, Eng., Izabela Koss-Mikołajczyk, PhD, Eng., and Monika Kuczyńska, PhD, Eng. won the distinction of the Division II of Biological and Agricultural Sciences of the Polish Academy of Sciences.
This year, five Divisions of the Polish Academy of Sciences awarded researchers for outstanding scientific achievements. Individual scientists and research groups were appreciated.
Every year, the Academy's Divisions award prizes for outstanding achievements and outstanding scientific papers. They are received by researchers employed in Polish units. Candidates are proposed by the Discipline Councils of parent scientific units, and recommended by members of the Academy and research and problem-oriented committees. The winners and laureates are selected by each of the PAN divisions.
The research team from the Faculty of Chemistry of Gdańsk University of Technology, composed of: prof. Agnieszka Bartoszek-Pączkowska, PhD, DSc, Eng., Izabela Koss-Mikołajczyk, PhD, Eng., and Monika Kuczyńska, PhD, Eng. was awarded for an outstanding research achievement aimed at explaining the observation that food of plant origin containing bioactive antioxidant phytochemicals has a better health-promoting effect than those substances isolated from plants.
Studies of bioactive antioxidant phytochemicals conducted for several years have shown that the plant matrix and other bioactive compounds present in plants affect the final biological activity. This means that consuming dietary supplements containing compounds isolated from fruits or vegetables does not bring the same benefits to our body as eating those fruits or vegetables from which the substances were extracted.
Thus, commonly used dietary supplements containing antioxidant compounds isolated from plants, being foodstuffs for which documentation is not required (as in the case of drugs), may not show the results declared by the manufacturers, proposed on the basis of studies described in the scientific literature and conducted for individual substances. The team's key achievement is proving that plant-derived substances act differently in different configurations, which makes it impossible to predict the effects of specific ingredient systems without medical research.
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Artificial intelligence achieves near-human performance in breast cancer diagnosis
Pathologists have been diagnosing disease the same way for the past 100 years, by manually reviewing images under a microscope. But new work suggests that computers can help doctors improve accuracy and significantly change how cancer and other diseases are diagnosed
Pathologists have been largely diagnosing disease the same way for the past 100 years, by manually reviewing images under a microscope. But new work suggests that computers can help doctors improve accuracy and significantly change the way that cancer and other diseases are diagnosed.
A research team from Beth Israel Deaconess Medical Center and Harvard Medical School, both in Boston, Massachusetts, recently developed artificial intelligence (AI) methods aimed at training computers to interpret pathology images, with the long-term goal of building AI-powered systems to make pathologic diagnoses more accurate.
"Our AI method is based on deep learning, a machine-learning algorithm used for a range of applications including speech recognition and image recognition," explained pathologist Andrew Beck, MD, PhD, Director of Bioinformatics at the Cancer Research Institute at Beth Israel Deaconess Medical Center and an Associate Professor at Harvard Medical School. "This approach teaches machines to interpret the complex patterns and structure observed in real-life data by building multi-layer artificial neural networks, in a process which is thought to show similarities with the learning process that occurs in layers of neurons in the brain's neocortex, the region where thinking occurs."
The Beck laboratory's approach was recently put to the test in a competition held at the annual meeting of the International Symposium of Biomedical Imaging (ISBI), which involved examining images of lymph nodes to decide whether or not they contained breast cancer. The research team of Beck and his lab's post-doctoral fellows Dayong Wang, PhD and Humayun Irshad, PhD, and student Rishab Gargya, together with Aditya Khosla of the MIT Computer Science and Artificial Intelligence Laboratory, placed first in two separate categories, competing against private companies and academic research institutions from around the world.
The research team recently posted a technical report describing their approach to the arXiv.org repository, an open access archive of e-prints in physics, mathematics, computer science, quantitative biology, quantitative finance and statistics.
"Identifying the presence or absence of metastatic cancer in a patient's lymph nodes is a routine and critically important task for pathologists," Beck explained. "Peering into the microscope to sift through millions of normal cells to identify just a few malignant cells can prove extremely laborious using conventional methods. We thought this was a task that the computer could be quite good at – and that proved to be the case."
Automated breast cancer diagnosis
In an objective evaluation in which researchers were given slides of lymph node cells and asked to determine whether or not they contained cancer, the team's automated diagnostic method proved accurate approximately 92 per cent of the time, explained Khosla, adding, "This nearly matched the success rate of a human pathologist, whose results were 96 per cent accurate."
"But the truly exciting thing was when we combined the pathologist's analysis with our automated computational diagnostic method, the result improved to 99.5 per cent accuracy," said Beck. "Combining these two methods yielded a major reduction in errors."
The team trained the computer to distinguish between cancerous tumour regions and normal regions based on a deep multilayer convolutional network. "In our approach, we started with hundreds of training slides for which a pathologist has labeled regions of cancer and regions of normal cells," said Wang. "We then extracted millions of these small training examples and used deep learning to build a computational model to classify them."
The team then identified the specific training examples for which the computer is prone to making mistakes and re-trained the computer using greater numbers of the more difficult training examples. In this way, the computer's performance continued to improve.
"There have been many reasons to think that digitising images and using machine learning could help pathologists be faster, more accurate and make more accurate diagnoses for patients," Beck added. "This has been a big mission in the field of pathology for more than 30 years. But it's been only recently that improved scanning, storage, processing and algorithms have made it possible to pursue this mission effectively.
"Our results in the ISBI competition show that what the computer is doing is genuinely intelligent and that the combination of human and computer interpretations will result in more precise and more clinically valuable diagnoses to guide treatment decisions."
Jeroen van der Laak, PhD, who leads a digital pathology research group at Radboud University Medical Center in the Netherlands and was an organiser for the competition, said, "When we started this challenge, we expected some interesting results. The fact that computers had almost comparable performance to humans is way beyond what I had anticipated. It is a clear indication that artificial intelligence is going to shape the way we deal with histopathological images in the years to come."
Beck and Khosla recently formed a start-up company (PathAI) with the mission of developing and applying AI technology for pathology.
https://www.engineersjournal.ie/2016/07/12/artificial-intelligence-achieves-near-human-performance-breast-cancer-diagnosis/https://www.engineersjournal.ie/wp-content/uploads/2016/07/breast-cancer-diagnosis-1024x683.jpghttps://www.engineersjournal.ie/wp-content/uploads/2016/07/breast-cancer-diagnosis-300x300.jpg 2016-07-11T21:59:42+00:00 David O'RiordanBiobiomedical,cancer,research
Pathologists have been largely diagnosing disease the same way for the past 100 years, by manually reviewing images under a microscope. But new work suggests that computers can help doctors improve accuracy and significantly change the way that cancer and other diseases are diagnosed. A research team from Beth Israel...
David O'Riordandoriordan@engineersireland.ieAdministratorEngineers Journal
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Bionic pancreas controls blood-sugar levels in type 1 diabetes The latest version of a bionic pancreas device has...
Prototype, pilot and production – outsourcing manufacturing for medical devices Author: Bill Peatman, CIRTEC Medical Systems Medical device manufacturing...
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Mystic, CT - Humpback whales and walruses now at Mystic Aquarium? Not literally but virtually, beginning this weekend. Just in time for school vacation, Mystic Aquarium will debuted Wild Arctic, an augmented reality journey that will provide guests an unbelievable encounter with some of the Arctic's most amazing creatures.
"We believe this hi-tech exhibit is one more way that we can help establish a deeper connection with our visitors to this remote environment and its engaging inhabitants," said Katie Cubina, Senior Vice President, Mission Programs.
Of course, the most compelling way that Mystic Aquarium engages children and families is through important animal ambassadors like its beluga whales.
Wild Arctic uses multiple high-end projection systems and state-of-the-art computer-generated animation to completely transform the exhibit area into a fully immersive trip to the Arctic. Through the magic of augmented reality, visitors come face-to-face with a live streaming video of themselves interacting virtually with wild animals.
Visitors are immediately swept up into an encounter with Arctic species including a polar bear, walruses and a breaching humpback whale. By stepping onto the 'hot spot,' guests are completely immersed into the Arctic adventure.
The addition of the Wild Arctic exhibit further solidifies Mystic Aquarium as a destination for interactive family fun.
But it's not all just fun and games.
A rapidly changing Arctic poses concerns to the welfare of marine mammals in the natural environment. By creating authentic connections through exhibits like the Arctic Coast Beluga Whale habitat and simulations like the Wild Arctic Augmented Journey, Mystic Aquarium is able to reach mass audiences to inspire a greater understanding of this incredible region and promote a conservation ethic and action for change.
While visitors engage with augmented reality in Wild Arctic, Mystic Aquarium scientists and veterinarians are actively engaged in pure and applied research aimed at conserving Arctic species.
Each year, Mystic Aquarium scientists conduct research expeditions to Alaska and other Arctic locations to monitor the health of wild populations of beluga whales and determine whether climate change, pollution and other factors are having an impact on their well-being. This work can and will directly impact the Arctic ecosystem and its inhabitants like beluga whales, polar bears, walrus and others.
Mystic Aquarium continually strives to create new ways, like through the use of Augmented Reality showcased in Wild Arctic, to build the connection, channel the social action and promote positive conservation behaviors.
"It's the Arctic within your reach, right here in Connecticut," added Cubina.


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Q: Permanently Delete the sql server database table I wanted the remove the table from the sql server database. The table has 500 million records. I don't need the table anymore.
What's the best approach?
Can I use drop table command to delete the table permanently? If yes is this the good approach?
I want to minimize the transaction log file growth or any other issues while I drop the table.
Should I do in off business hours or if the impact is minimal I can it anytime?
SQL server version is both 2017 and 2019.
Databases containing those tables have same compatibility and are in full recovery model.

A: DROP TABLE is a minimally logged operation, as already answered here.
So, you can go ahead with the DROP TABLE operation.

redpajama_set_name: RedPajamaStackExchange


A gorgeous piece of art for serving or display—solid blocks of wood are sourced from fallen trees in Western Massachusetts and hand-turned on a lathe, making each bowl totally unique. Ambrosia maple is prized for its beautiful and distinctive coloration, caused by the ambrosia beetle. The bowls are oiled and coated in a food-safe beeswax to protect them from drying out. Available in three sizes; the 9" and 11" are great for side dishes, and the 13" is the perfect salad bowl.
Spencer Peterman has been hand-turning wooden bowls in his workshop in Gill, Massachusetts since 2000. Each bowl is signed by the artist.


Nespresso machines are automatic espresso machines made by Nespresso. They brew espresso of various sizes simply at the push of a button. The machines run on specially packaged "pods" or "capsules" that contain the coffee perfectly ground to work with the machines. The pods of course come in many "flavors" and types of beans and roasting darknesses.
The pods also come in consumer style and professional style. The consumer version is shaped and sized like a thimble whereas the pro version is shaped more like a UFO in the size of a Snapple cap. Consumer pods are meant for consumer machines and Pro pods are meant for the Pro line of machines. The two cannot be used interchangeably.
Nespresso recommends using filtered or distilled water. Experience says this is good advice as long usage with tap water can eventually develop deposits and scaling on the machine. Continued usage as such may render the machine inoperable or at the least in need of service.


 T. Mert , Gunes Y, Gunay I Comparison of actions of systemically and locally administrated local anaesthetics in diabetic rats with painful neuropathy. Fundam Clin Pharmacol. 2011 Oct 11. doi: 10.1111/j.1472-8206.2011.00998.x. [In Press].
 T. Mert , Y Gunes, D Ozcengiz. Modification of local anesthetic-induced antinociception by fentanyl in rats. Pharmacological Reports 63: 1427-1434, (2011).
 Gunay, I.ve T. Mert. "Pulsed magnetic fields enhance the rate of recovery of damaged nerve excitability." Bioelectromagnetics 32(3): 200-208 (2011).
 T Mert, I Gunay, I Ocal. "Neurobiological Effects of Pulsed Magnetic Field on Diabetes Induced Neuropathy." Bioelectromagnetics 31, 39-47 (2010).
 T. Mert, Gunes, Y., Ozcengiz D ve Gunay, I. "Magnesium modifies fentanyl-induced local antinociception and hyperalgesia." Naunyn-Schmied. Arch. Pharmacol 380, 415-420 (2009).
 T. Mert, Gunay, I., Ocal I., Guzel AI., Inal TC., Sencar L. ve Polat S. "Macrophage depletion delays progression of neuropathic pain in diabetic animals". Naunyn-Schmied. Arch. Pharmacol. 379, 445-452 (2009).
 T. Mert, Gunes, Y., Gunay, I. "Role of 4-Aminopyridine-Sensitive Potassium Channels in Peripheral Antinociception". Eur. J. Pharmacol. 572, 138-141 (2007).
 T. Mert, "Roles of Axonal Voltage-Dependent Ion Channels in Damaged Peripheral Nerves." Eur. J. Pharmacol. 568, 25-30 (2007).
 T. Mert, "Sucrose-Gap Technıque: Advantages and Lımıtatıons". Neurophysiology 39(3), 270-274 (2007).
 T. Mert, Gunes, Y., Gunay, I. "Local Analgesic Efficacy of Tramadol Following Intraplantar Injection". Eur. J. Pharmacol. 558, 68-72 (2007).
 T. Mert, Gunay, I., Gocmen, C., Kaya, M., Polat, S. "Regenerative Effects of Pulsed Magnetic Field on Injured Peripheral Nerves". Alter. Ther. Health Med. 12(5), 42-49 (2006).
 T. Mert, Gunes, Y., Ozcengiz, D., Gunay, I.. Polat, S. "Comparative Effects of Lidocaine and Tramadol on Injured Peripheral Nerves" Eur. J. Pharmacol. 543, 54-62 (2006).
 T. Mert, "Kv1 Channels in Signal Conduction of Myelinated Nerve Fibers" Rev. Neurosci. 17(3), 369-373 (2006).
 T. Mert, Günay, I. Polat, S. "Alterations in Conduction Characteristics of Crushed Peripheral Nerves". Rest. Neurol and Neurosci 23(5-6), 347-354 (2005).
 Guven, M., T. Mert, Günay, I. "Effect of Tramadol on Nerve Action Potentials: Comparisons with Lidocaine and Benzocaine" Int J Neurosci. 115(3), 339–349 (2005).
 T. Mert, Daglioglu, Y.K., Gunay, I., Gocmen, C. "Changes in Electrophysiological Properties of Regenerating Rat Peripheral Nerves After Crush Injury". Neurosci Lett 363, 212-217 (2004).
 T. Mert, Gunes, Y., Guven, M., Gunay, I. Goçmen, C. "Differential Effects of Lidocaine and Tramadol on Modified Nerve Impulse by 4-Aminopyridine in Rat". Pharmacology, 69(2), 68-73 (2003).
 T. Mert, Gunes, Y., Guven, M., Gunay, I. Ozcengiz, D. "Effects of Calcium and Magnesium on Peripheral Nevre Conduction". Pol J .Pharmacol 55, 25-30 (2003).
 T. Mert, Gunes, Y., Guven, M., Gunay, I., Ozcengiz, D. "Comparison of Nerve Conduction Blocks by An Opioid and A Local Anesthetic" Eur J Pharmacol, 439, 77-81, (2002).
 T. Mert, Gunes, Y., Guven, M., Gunay, I. Ozcengiz, D. "Blocking Action of Tramadol on Nerve Conduction" I J Pharm, 2(1), (2001).


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Asia News Network
US version of 'King of Masked Singer' hugely popular
By Yonhap
Published : Jan 27, 2019 - 10:41
Updated : Jan 27, 2019 - 10:41
An American version of the popular South Korean TV variety show "King of Masked Singer" has been a sensation in the United States, garnering over 10 million views per episode, the show's producers said Sunday.
The remade show's first episode was watched by 9.368 million when it premiered on the Fox channel on Jan. 2, and cumulative views, including runs on video on demand and over-the-top services, rose to 12.27 million and 12.96 million three and seven days later, respectively, according to Fox and MBC.
(Yonhap)
The second and third episodes each have also drawn more than 10 million views, they said.
MBC is the broadcaster of the original singing competition show.
In the US, programs attracting more than 10 million views are considered a big success. The cumulative views of "The Masked Singer" are more than the 6.91 million for a recent episode of the hit American television show "The Big Bang Theory."
"The Masked Singer" is one of three South Korean programs that have been remade in the US, along with tvN's "Grandpa Over Flowers" and KBS 2TV's "The Good Doctor."
The original Korean show "King of Masked Singer" premiered in April 2015 in Korea, and the format has been sold to about 20 countries, including Britain, France, Thailand, Vietnam, Indonesia and China.
The program is still in service in South Korea, boasting a strong viewership hovering around 10 percent. (Yonhap)


A variety of mechanized welding systems are available from Lincoln Electric including welding booms. We offer a range of manipulators and automated welding systems that can perform operations including MIG, FCAW, and submerged arc welding. These systems are perfect for handling repetitive welding operations as they will increase productivity and speed to market. Whether you require a welding boom or a complete automated system, we can provide an appropriate solution from our wide selection. Our product offering also includes orbital systems, cutting equipment, training equipment, and weld fume control.
Our welding manipulator systems are designed to perform various combinations of welding processes. They feature the ability to handle heavy loads with smooth, consistent movement on all axes. These manipulators can be either floor mounted using a free stand base or mounted to a travel car. The boom and hoist movement, variable speed control, slides and travel car can all be controlled with the remote pendant.
The manipulators are available in different sizes and configurations. They can offer horizontal boom travel up to 3.4 meters (11 feet) at variable speeds from 3 to 116 inches per minute. Manipulators are supplied with a digital speed indicator, over travel limit switch, single wire and tandem SAW mounting kits, and more.
In addition to welding booms and manipulators, we can provide complete, integrated solutions that include slides, positioners, controllers, power source, and more. Our mechanized welding systems can be customized to suit the application. We have the experience and capabilities to create solutions that make our customers more productive and successful.
As a global manufacturer and market leader, we are known for the high quality of our welding, cutting, and joining products. Lincoln Electric has 120 years of experience serving a variety of industries including automotive, offshore, power generation, mining, and agriculture. To meet customers' unique requirements, we can offer specialized materials, product formulations, quality assurance programs, and packaging.


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Global Control, Rothschild Bacharach March 10, 2022 March 10, 2022
General Electric (GE) is a Rothschild-led and -controlled (J.P. Morgan) company, from inception. Therefore, RCA/NBC offspring were and still are Lords Rothschild controlled.
GE's Coffin and Griffin were founding members of the British Pilgrims Society in 1902.
Editors. (Sep. 01, 1977). The Steinmetz Era, 1892-1923, The General Electric Story, Vol. 2, A Photo History. Elfun Hall of History.
https://www.fbcoverup.com/docs/library/1977-09-01-The-Steinmetz-Era-1892-1923-The-General-Electric-Story-Vol-2-A-Photo-History-Elfun-Hall-of-History-Sep-01-1977.pdf#page=8
J.P. Morgan had just inherited his Rothschild-funded bank from his dead father Junius in 1890, just two years earlier.
Re: Charles A. Coffin, GE President
https://en.wikipedia.org/wiki/Owen_D._Young formed RCA as a part of GE in 1919 and was its first president and chairman until 1929.
Marconi Wireless operator David Sarnoff, founder of NBC (1925), succeeded Owen as president of RCA (1929) when Owen Young retired.
Gerald Swope GE President (1922-40)
J.P. Morgan in 1892 was a notorious agent for Lord Lionel Rothschild. Just two years earlier, Morgan inherited Junius Morgan & Sons Bank who was the successor of George Peabody banks bankrolled by N.M. Rothschild.
Choice find from a state library archivist – this New York Times article appeared just nine months after the Pilgrims Society organized the day after the enthronement of Edward VII on Aug. 09, 1902.
Notice that General Electric joined immediately. (to our knowledge this is the first evidence about GE's foundational involvement in the UK)
Editor. (May 17, 1903). THE PIGRIMS' SOCIETY GROWS, More Notable American and Englishmen Join Organization. The New York Times. Reproduced for educational purposes only. Fair Use relied upon. Source: https://www.nytimes.com/1903/05/17/archives/pilgrims-society-growsmore-notable-americans-and-englishmen-join.html
Also published in Indiana:
Indianapolis Journal, Volume 53, Number 137, Indianapolis, Marion County, 17 May 1903
https://newspapers.library.in.gov/cgi-bin/indiana?a=d&d=IJ19030517.1.5&e=——-en-20–1–txt-txIN——-
More links of interest:
https://circleofthedolphins.wordpress.com/tag/rockefeller-rothschild/
https://brutalproof.net/2021/04/very-important-information-about-the-crime-syndicate-running-america-pilgrims-society/
http://eurofolkradio.com/2019/09/01/voci-the-pilgrims-society-exposed-as-a-rothschild-front/


Umberto Bonomo is a young producer and promotor of architecture, a participant of the currently most vibrant architecture scene, that of Chile.
He is an Urbanist and Architect by IUAV (Venice, 2004) and PhD in Architecture and Urban Studies by Catholic University (Santiago, 2009). Is Sub-Director in Development area and Assistant Professor in The School of Architecture at Pontificia Universidad Católica in Santiago. Is a researcher in Urban and Architecture Theory, Housing Projects and Heritage. He works as a designer in his own practice. His projects and researches have been published and exposed in many books, reviews and lectures, such as "Insularity and Difference" Columbia University NY (2009), Venice Biennale (2010), Chilean Biennale (2010), Expo Shanghai (2010), BIAU (2012), Sao Paulo Bienal (2013), Shenzhen Biennale (2016).
Umberto Bonomo held an interview with Alejandro Aravena which you can rear in Oris 100.


Are you troubled by visions? - Knowlan Consulting Group Inc. | Knowlan Consulting Group Inc.
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Several of our clients have confided recently that they are troubled by "visions". But before you reach for that long-sleeved canvas jacket, note that they're talking about vision statements! Common concerns we've heard include whether having a vision statement is important or, for those who already have one, whether it's actually helping.
Almost any organization would benefit with the introduction of a clear, compelling vision that convincingly summarizes leaders' aspirations and infuses employees with a renewed sense of purpose. Creating a vision statement that raises everyone's expectations and performance is a tall order. But if your vision statement doesn't elevate commitment and effort to higher levels, what good is it?
After helping many organizations to create vision statements, we're struck by how carefully the process must be managed to keep it on track. Many visioning exercises consume inordinate amounts of time only to fall far short of sparking better performance. In this article, we'll explain the most common problems we've encountered in visioning – problems with timing, involvement, and format – and offer tips to help you avoid them.
Starting a visioning exercise at the wrong time is almost a sure-fire recipe for failure. If you've tried visioning while overloaded or in crisis, you'll appreciate how hard it is to "dream big" when you're living a nightmare! As one manager in a troubled organization said about senior management's new vision, "It looks like a wonderful place, but we can't get there from here".
The timing of a visioning process matters, even when things are going smoothly. Many clients ask to add visioning to their strategic planning agenda, but we recommend against it. Strategic planning is a rigorous, logical and analytical process. Being realistic is often participants' highest planning priority since each of them will carry responsibility for delivering results on a time-line. Visioning, on the other hand, is a creative, expansive, optimistic and dream-nurturing process. The rigorous realism of strategic planning is a poor fit with the expansive optimism of visioning, and most teams find it difficult to shift between these mind-sets in the same meeting. When strategic planning and visioning are attempted concurrently, one of them is likely to suffer.
Finally, your senior team will strongly influence the outcome of a visioning exercise. If they are satisfied or complacent about current performance, the status quo is the de facto vision, no matter how ambitious other participants may be! Unless your senior team is strongly motivated to create a brighter future for the organization, visioning is likely to be a pointless exercise.
It's important to build support for a new vision by drawing input from across the organization, but be careful to involve suitable people in the right roles. The part of your vision statement devoted to values is a good subject for input from a broad range of organizational members. But be selective when deciding whom to involve in setting a visionary goal. This goal must embody boldness as well as believability before it will move people to make significant changes. The ideas for this must come from your visioning team – their challenge will be to blend ambitious ideas with astute judgment about what is possible far in the future.
Visioning requires at least a few participants who are both visionary and judicious! Make sure your team has the right people to meet this challenge. While many aspects of visioning require broad involvement, writing the final statement is not one of them! If you expect your vision statement to inspire people in ways that stretch their expectations, aspirations and performance, it must be artfully written – a task that is beyond the capability of virtually any large team. Excellent writing skills are uncommon, but are essential for effective visioning. The visioning team should include at least one writer who can meld the team's ideas into succinct and eloquent language.
While most teams readily agree on the criteria for common terms like "objectives", we've seen visioning processes stumble over differing views about the "right" format and content of the resulting statement. For example, one team got deadlocked over whether their vision should be "short and punchy" like Nike's just do it(1) slogan, or a longer, content-rich version like the Built to Last formula(2). This is not a trivial choice, and there's no "correct" length. A long vision won't inspire anyone who finds it boring, but if it's too short, it won't provide a clear purpose and direction.
Another common conundrum for visioning is choosing the time horizon. Many feel more comfortable creating a vision that is intended to be reached in 5 to 7 years; others advocate a much longer view, arguing it is impossible to get airborne from a runway that is too short. Selecting a shorter time horizon tends to reduce people's willingness to commit to ambitious undertakings. For example, we wonder whether Bill Gates would have aspired in 1975 to equip every business and household with a computer if it had to be achieved in only 5 to 7 years. It is generally better to begin a visioning process with a longer time horizon in mind to avoid imposing an arbitrary constraint on the outcome. If the visioning team sets a goal that is achievable in a shorter period, the time horizon can be shortened accordingly.
Without prior agreement on the length, content and time horizon of the statement, a visioning exercise will probably take longer and produce an inferior outcome.
Follow these tips to increase the likelihood of success in creating or modifying your vision statement.
• Choose your timing well – not during a crisis or overload, and not during strategic planning. Teams don't usually agree on ambitious visions unless they're feeling confident and successful, and combining visioning with analytical tasks is likely to stifle optimism and creative thinking.
• Choose carefully who will be involved at which steps. You'll need broad involvement, which is relatively easy to arrange, and visionary thinking, which is not. And you'll need one or two people with strong writing skills to convert ideas into coherent statements and polish drafts between meetings.
• Make sure you have a good reason to create a vision statement. If the executive team doesn't feel a gap between their aspirations and current reality, they aren't ready for visioning. And you may not need an "official" vision statement if your strategic plan already has ambitious growth built into it. A commitment to become the dominant player in your competitive space, for example, may keep your company focused and striving for many years.
• Don't start without first setting format guidelines including the length, content and time horizon of your statement. You may decide to vary from these guidelines as you proceed, but stick with your original format until you're convinced a change is needed.
• Our clients become skilled at updating and evolving strategic plans, but the same can't be said for visioning. Most undertake visioning so seldom that they underestimate how different the two processes are. The hard-headed realism that serves strategic planning so well generally makes visioning far too conservative. If you're about to embark on a visioning process to create a bright new future for your organization, consider retaining external help to maximize your chances that your new vision will raise everyone's expectations and performance.
Embarking on a visioning exercise does not guarantee success, and since it raises people's expectations, failure to reach a satisfying conclusion is likely to be discouraging to everyone involved. Choosing your timing, format and participants wisely will help you achieve a better result.
© 2007 Knowlan Consulting Group Inc. All rights reserved. Unauthorized duplication or publication in any form, in whole or in part, is prohibited.
1. Nike's "Just Do It" was never intended as a vision statement.
2. "Built to Last: Successful Habits of Visionary Companies"; James C.
Collins, Jerry I. Porras, 1994.
At Power Smart we hit upon the the thought "to create a world where energy efficiency is a way of life". . . In other words people just did it without much thought because of course that's what we do. Amory Lovins the efficiency Guru remarked that we were the first utility to actually define a vision. Maybe that's what made us better.
I often use the Power Smart Mission with its two parts as an example of an elegant Mission and Vision. It encompasses the immediate payback for the customer (best value for their uses of electricity) and the bigger value for society (create a world where energy efficiency is a way of life).


The Bloody Crossroads is a website about politics and culture. It takes its name from the critic Lionel Trilling, who wrote about 'the dark and bloody crossroads where literature and politics meet'. By that he meant the way in which literature is inseparable from our view of the world; but he might also have meant the way in which politics is inseparable from the way we talk. If there is a common theme in the essays and articles collected at The Bloody Crossroads, this is it: a concern with the politics of poetry and the poetry of politics.
There are other crossroads, just as dark and bloody, explored in these pages. One recurrent theme – familiar to Trilling and his fellow New York intellectuals – is the tension between, and the effort to reconcile, two distinct political traditions: liberalism and socialism, or a concern for individual rights and a desire for a more inclusive, equal and egalitarian society. This may sound like an arcane consideration, but it bears on many of our current debates, from the limits of free speech to the status of religion. All of the writers I most admire were concerned with this fundamental question.
Finally, there is the crossroads at which the world finds itself at the present moment. Humankind stands at a crucial juncture, where climate change, gross and growing inequality and a collapsing economic system threaten (promise) to remake the world. One consequence of this is that ideas are big again, and if these articles do nothing else they may help to ventilate a few them. I hope so, and I hope that visitors to The Bloody Crossroads will find direction, even if they don't find answers.