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"Q:\n\nAndroidAnnotations. ",
"button click inside actionLayout of menuItem\n\nIn AA+AppCompat, I try to change below code in Activity to AA-style.",
"\n@Override\npublic boolean onCreateOptionsMenu(Menu menu) {\n MenuInflater menuInflater = getMenuInflater();\n menuInflater.inflate(R.menu.menu_done, menu);\n MenuItem menuItem = menu.findItem(R.id.itemDone);\n View menuView = MenuItemCompat.getActionView(menuItem);\n mButtonQuestionPost = (Button) menuView.findViewById(R.id.buttonMenuDone);\n mButtonQuestionPost.setOnClickListener(this);\n\n return super.onCreateOptionsMenu(menu);\n}\n\nFirst, changed head of Activity as below. ",
"The menu was shown as expected.",
"\n@EActivity(R.layout.activity_question_post)\n@OptionsMenu(R.menu.menu_done)\npublic class QuestionPostActivity extends FragmentActivity {...\n\nAnd, try to do button click method as below but nothing fired. ",
"I changed @Click with @OptionsItem or changed attributes this and that but no luck.",
"\n@Click(R.id.buttonMenuDone)\nvoid buttonMenuDone(){\n if (mQuestionPostFragment.validatePost()) {\n setSupportProgressBarIndeterminate(true);\n mQuestionPostFragment.postQuestion();\n }\n}\n\nmenu.xml is as below. ",
"Note that I'm using actionLayout for design purpose.",
"\n\n<item\n android:id=\"@+id/itemDone\"\n android:title=\"@string/done\"\n app:showAsAction=\"always\"\n android:menuCategory=\"system\"\n app:actionLayout=\"@layout/item_menu_done\"\n />\n\nitem_menu_done is as below.",
"\n\n<Button\n android:layout_width=\"48dp\"\n android:layout_height=\"?actionBarSize\"\n android:id=\"@+id/buttonMenuDone\"\n android:text=\"@string/done\"\n android:textColor=\"@android:color/white\"\n android:textSize=\"14sp\"\n android:background=\"?attr/actionBarItemBackground\"\n />\n\nThe AA-generated file does not have MenuItemCompat.getActionView(menuItem), but hardly to make it with AA. ",
"Can someone please help me?",
"\n\nA:\n\nI am afraid you cannot bind a listener with @Click to a menu action view, since Activity.findViewById cannot find that view inside the menu items. ",
"What you can do is injecting the menuitem, then, manually bind your listener as you already did.",
"\n@EActivity(R.layout.activity_question_post)\n@OptionsMenu(R.menu.menu_done)\npublic class QuestionPostActivity extends FragmentActivity {\n\n @OptionsMenuItem(R.id.menuItemDone)\n MenuItem buttonMenuDone;\n\n @Override\n public boolean onCreateOptionsMenu(Menu menu) {\n // this will be called by the generated subclass after it injected the menu\n MenuItemCompat.getActionView(itemDone).findViewById(R.id.buttonMenuDone).setOnClickListener(this);\n return true;\n }\n}\n\n"
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"ISO/IEC TR 12182\n\nISO/IEC TR 12182 is an Information technology standard published in 1998 by the Joint Task Committee 1 (JTC1) of the International Organization for Standardization (ISO) and the International Electrotechnical Commission (IEC). ",
"It defines a software categorization in the field of software engineering.",
"\n\nReferences \n International Organization for Standardization\n International Electrotechnical Commission\n JTC1\n Veridion ISO 27001 Directory\n ISO 12182 Standard\n\n#12182"
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[
"Driven dynamics of disordered systems have been studied extensively from two different approaches. ",
"As the external driving force is increased, a system undergoes a depinning transition from a macroscopically static state (with transient motion) to a moving steady state. ",
"This transition has been successfully described as a dynamical second order phase transition [@DSF], and analyzed within the framework of the renormalization group (RG), originally for charge-density waves [@CDW] and subsequently for interfaces [@interf] and a variety of other systems [@graball; @ONAM]. ",
"This approach obtains the scaling behavior near the transition where the system starts to move. ",
"On the other hand, the concept of self-organized criticality (SOC), in the original sandpile model [@BTW] and descendants thereof [@PMB] has been used to obtain the behavior of systems that are forced to stay [*at*]{} the depinning transition.",
"\n\nOne would expect there to be connections between these two viewpoints [@Tang]; indeed, it is possible to map automaton models for CDWs to the original SOC model [@ONAM; @foot0], and 1+1 dimensional moving interface to a slightly different SOC model [@Pac1]. ",
"The connections between the two have been exploited to obtain the dynamical critical exponent $z$ for CDWs by borrowing from exact results for SOC sandpiles [@MD]. ",
"However, it has been pointed out [@PMB] that one has to be careful whether the depinning transition is approached with a time independent external driving force, tuned to its critical value, or with a time independent (infinitesimal) velocity. ",
"In the latter case, one envisages a feedback loop that adjusts the external force in a time-dependent way, so as to ensure that the rate at which the system moves (averaged over its entire spatial extent) is strictly time independent. ",
"It has been suggested [@PMB] that the critical behavior at the depinning transition could be different for these two driving modes. ",
"As an extreme example of constant current driving, there is the class of “extremal models\" [@Sneppen; @LesTang; @TGR], where at every (discrete) time step, there is activity only at one lattice site in the system.",
"\n\nFor constant force driving, there are fluctuations in the (spatially averaged) velocity of the system, while for constant velocity driving there are fluctuations in the external force. ",
"In either case, at a non-zero driving rate, the fluctuations are small for sufficiently large systems, and thus the two driving mechanisms should be equivalent. ",
"However, the large system limit is problematic at the critical point. ",
"For instance, with constant force driving, the mean square fluctuations in the velocity of a large system are $(\\delta v)^2\\sim\nv^2 (\\xi/L)^d, $ where $v$ is the mean velocity, $\\xi$ is the correlation length at velocity $v$ (defined through the velocity autocorrelation function or other methods), $L$ is the linear extent of the system, and $d$ is its dimensionality. ",
"One would expect that holding the velocity constant by adjusting the force will make a qualitative difference to the dynamics when the velocity fluctuations are a significant fraction of the mean velocity, [[*i.e.*]{} ]{}$\\xi\\sim L.$ Heuristically, for a fixed $v$ and $L\\rightarrow\\infty,$ imposing a constant velocity does not affect the dynamics, since the total ‘activity’ at every time step is $v L^d,$ so that avalanches etc. ",
"are free to unfold with their own internal timescales. ",
"On the other hand, for fixed $L$ as $v\\rightarrow 0,$ imposing a constant velocity ‘chokes off’ the dynamics: avalanches proceed sequentially, with internal dynamics determined by the velocity constraint.",
"\n\nIn this paper, we show that (to the extent that continuum descriptions are appropriate) both constant force and constant velocity driving actually have the [*same*]{} scaling under the renormalization group. ",
"However, scaling functions for various quantities, which can have different behavior in the limits $\\xi << L$ and $\\xi >> L, $ are often different in the $\\xi >> L$ regime for the two driving modes. ",
"It is standard for scaling functions in equilibrium critical phenomena to behave differently when their arguments are small and large, but as will be discussed later, dynamical critical points have even more freedom in how scaling functions can behave. ",
"It is this freedom that allows identical RG flows to still yield different scaling functions for the two driving modes in the $\\xi >> L $ regime.",
"\n\nWe demonstrate this by focusing on a particular system, that of a interface with short range internal elastic interactions that is driven through a disordered medium under the influence of an external force. ",
"Due to the disorder, different parts of the interface experience different random pinning forces as they move forward. ",
"Using the RG relationship between the interface velocity and the correlation length [@interf] $v\\sim\\xi^{\\zeta - z},$ we see that the crossover to $\\xi> L$ occurs at $v\\sim L^{\\zeta - z}.$ Note that extremal dynamics for a $d+1$ dimensional interface corresponds to $v\\sim L^{-d},$ and $z - \\zeta < d$ in all dimensions, so that $\\xi >> L$ for large $L$.\n\nIt has been pointed out [@SnepJen; @LesTang; @KTR] that in extremal models for the motion of pinned interfaces, the roughness of the interface can scale anomalously. ",
"Thus for an 1+1 dimensional interface in a system of size $L$ in steady state [@foot4], if $h(x, t)$ is the interface position as a function of the transverse coordinate $x$ and the time $t$, the roughness $w(t)=[\\langle\\{h(x, t + t_0) - \\overline h(t + t_0) - h(x, t_0) + \n\\overline h(t_0)\\}^2\\rangle]^{1/2}$ (all averages are spatial averages) has the scaling form [@KTR; @foot1] $$w(t) = t^\\beta_v L^{- 1/2} \\varphi\\Bigg({L\\over\n{t^{1/z_v}}}\\Bigg).",
"\n\\label{scalform}$$ (The subscripts on $\\beta$ and $z$ denote constant velocity driving.) ",
"It can be shown [@LesTang; @KTR] that the scaling function $\\varphi$ goes to a constant for large values of its argument. ",
"The explicit $L$ dependence that remains is anomalous, contrary to the normal expectation of a well defined $L\\rightarrow\\infty$ limit. ",
"As a consequence of this, if the long time (steady state) roughness scales as $w(t\\rightarrow\\infty)\\sim L^\\zeta,$ then the conventional relation $\\zeta = z \\beta$ is replaced with $$\\zeta = z_v\\beta_v - {{1\\over 2}}.",
"\n\\label{roughexp}$$ With the additional result [@LesTang; @KTR; @foot2] $$z_v = 1 + \\zeta\n\\label{dynexp}$$ one can obtain $\\beta_v$ in terms of $\\zeta.$ Both Eq.(\\[roughexp\\]) and Eq.(\\[dynexp\\]) differ from renormalization group (RG) results [@interf] for an interface driven with a constant applied force. ",
"In particular, the RG analysis yields [*no*]{} simple relation between the dynamic exponent $z$ and the roughness exponent $\\zeta,$ unlike Eq.(\\[dynexp\\]). ",
"Nor is there any reason to expect the anomalous scaling form of Eq.(\\[scalform\\]), and therefore $\\zeta=z\\beta$ should hold instead of Eq.(\\[roughexp\\]).",
"\n\nWe first demonstrate that, despite appearances, the RG flows are unaffected by going from a constant force driving mode to a constant velocity driving mode. ",
"The standard equation for zero-temperature driven dynamics of an interface is [@KLT] $$\\partial_t h(x, t) = \\nabla^2 h(x, t) + Y(h(x, t); x) + F.\n\\label{FLR}$$ Here $\\nabla^2 h(x, t)$ comes from the (short-ranged) elastic energy of the interface, $Y(h; x)$ is a pinning force that comes from a random (impurity) potential, and $F$ is the external driving force. (",
"Of course, it is not necessary that all lattice growth models —[[*e.g.*]{} ]{}the Sneppen model [@Sneppen]—can be mapped to this or any continuum equation.) ",
"We have neglected KPZ like terms [@KPZ]. ",
"Using the Martin Siggia Rose method [@MSR] and introducing an auxiliary field $\\hat h(x, t),$ one can construct [@interf] a generating functional $Z$ which can be written as $$\\begin{aligned}\nZ&=&\\int [dh][d\\hat h]\\exp\\Big[\\int\\!d^d x\\, dt \\nonumber\\\\\n& &\\qquad i\\hat h(x, t)\n\\{\\partial_t h - \\nabla^2 h - F - Y(h; x)\\}\\Big]\n\\label{msr1}\\end{aligned}$$ where integrating out the auxiliary field yields a product of $\\delta$-functions that imposes Eq.(\\[FLR\\]). ",
"If instead the interface is driven at constant velocity, Eq.(\\[msr1\\]) is replaced with $$\\begin{aligned}\nZ^\\prime&=&\\int [dF][d\\mu][dh][d\\hat h]\\exp\\Big[\\int\\!d^d x\\, dt \n\\nonumber\\\\\n& &\\qquad i\\hat h(x, t)\n\\{\\partial_t h - \\nabla^2 h - F(t) - Y(h; x)\\}\\Big] \\nonumber\\\\\n& &\\qquad\\qquad+ i \\mu(t)\\{v - \\partial_t h\\}.",
"\n\\label{msr2}\\end{aligned}$$ This extension is actually not difficult to understand: at any time $t,$ by first integrating over $\\mu(t)$ and $\\hat h(x, t), $ we obtain $\\delta$-function constraints that impose $\\langle\\partial_t h\n\\rangle = v$ (the average here is a spatial average) and Eq.(\\[msr1\\]) with a (as yet unknown) driving force $F(t).$ Now performing the integral over $F,$ the integral together with the first constraint sets $F(t)$ to be whatever it has to be for $\\langle \\partial_t h$ to be equal to $v.$ One is left with the second constraint, [[*i.e.*]{} ]{}Eq.(\\[FLR\\]), with $F(t)$ adjusted to ensure constant velocity. ",
"Even though $F(t)$ is now a dynamical variable instead of a parameter, since it is not a field, [[*i.e.*]{} ]{}it has no spatial dependence (nor has $\\mu(t)$), when short distance fluctuations are eliminated under renormalization there are no extra loop corrections in $Z^\\prime$ compared to $Z.$\n\nAs mentioned earlier, even though the RG fixed point is the same for constant force and constant velocity driving, the behavior of scaling functions are different for the two driving modes in the $\\xi >> L$ regime. ",
"To illustrate this, we first consider the scaling of the duration of an avalanche as a function of its linear size. ",
"This has the general scaling form $$t(l, L, \\xi) = l^z T(l/L, \\xi/L).",
"\n\\label{avaltime}$$ Here $l$ is the linear extent of the avalanche (in the $d$ transverse directions; in the direction the interface moves, the extent is $\\sim l^\\zeta$), $L$ is the linear size of the system, $\\xi$ is the correlation length, and $z$ is the non-trivial dynamical exponent from the RG. ",
"For a fixed $l$ and $\\xi,$ as $L\\rightarrow\\infty,$ the avalanche duration should be independent of $L$ since the system is uncorrelated on length scales much bigger than $\\xi$ [@foot3]. ",
"It should also be independent of $\\xi,$ since requiring a total growth rate of $v L^d$ for the system does not affect any individual avalanche from proceeding with its own intrinsic timescale. ",
"Thus $T(0, 0)$ is some non-zero constant.",
"\n\nIn the other regime of $\\xi>> L,$ [[*i.e.*]{} ]{}if the $v\\rightarrow 0$ limit is taken [*before*]{} $L\\rightarrow\\infty,$ the avalanches are non-overlapping in time. ",
"In the constant velocity driving mode, where the velocity constraint is imposed, any single avalanche proceeds at a fixed rate. ",
"Therefore for the constant velocity driving mode, $t(l, L, \\xi)$ must be inversely proportional to the rate at which the avalanche is allowed to proceed, which is $v L^d.$ Requiring that $t$ should have such a dependence on $v$ and $L,$ using the result [@interf] $v\\sim \\xi^{\\zeta - z},$ it is straightforward to verify that $$\\lim_{\\xi/L\\rightarrow\\infty} t(l, L, \\xi) \\sim \nl^{\\zeta + d }/ (v L^d).",
"\n\\label{avt}$$ This yields an apparent dynamical exponent of $z_v = \\zeta + d,$ which is the $d+1$ dimensional generalization of Eq.(\\[dynexp\\]). ",
"Note that there is [*no*]{} change in the apparent dynamical exponent in the $\\xi >> L$ regime for constant force driving, where an avalanche is allowed to proceed at its intrinsic rate (through parallel updating of lattice sites). ",
"This is why the RG estimate for $z$ for two dimensional CDWs agrees with the exact result for Abelian sandpiles [@MD].",
"\n\nThe analysis of the interface roughness for a system of linear size L, $w(t, L),$ proceeds in a similar manner. ",
"At a velocity $v,$ the interface roughness has the scaling form $$w(t, L, \\xi) = L^\\zeta W(t/L^z; \\xi/L).",
"\n\\label{roughscal}$$ For fixed $t$ and $\\xi$, the roughness must have a well defined $L\\rightarrow\\infty$ limit: $$\\lim_{L\\rightarrow\\infty} w(t, L, \\xi) = \\xi^\\zeta W_1(t/\\xi^z).$$ For large $t,$ the roughness saturates to the steady state form $w\\sim \\xi^\\zeta.$ For $\\xi\\rightarrow\\infty$ (the large $L$ limit has been taken first), or equivalently for small $t,$ the roughness is $\\xi$ independent, [[*i.e.*]{} ]{}$w\\sim t^{\\zeta/z}.$\n\nIn the other regime of $\\xi>> L,$ the apparent exponents are once again different for constant velocity driving. ",
"In this case, the dynamics are controlled by the imposed velocity, [[*i.e.*]{} ]{}the $v$ and $t$ dependence of Eq.(\\[roughscal\\]) occurs only in the combination $\\tau = vL^d t.$ Using $v\\sim\\xi^{\\zeta - z},$ this implies $$\\lim_{\\xi/L\\rightarrow\\infty} w(t, L, \\xi) = L^\\zeta W_2(\\tau/\nL^{\\zeta + d}).$$ (With extremal dynamics, where one site moves forward at every time step, $\\tau = t.$) In the large time limit, the roughness approaches the steady state form $w\\sim L^\\zeta, $ [[*i.e.*]{} ]{}$W_2(\\infty)$ is a constant. ",
"It is not obvious how to extract the behavior of the function $W_2$ when its argument is small, but physical arguments supported by numerical results [@LesTang; @KTR] show that $w(\\tau, L)$ must have a residual $L^{-d/2}$ dependence. ",
"Therefore $$w\\sim (v L^d t)^{\\beta_v}/L^{-d/2}\n\\label{rghsc}$$ with $$\\beta_v = {{\\zeta + d/2}\\over{\\zeta + d}}$$ which is the same as Eq.(10) of Ref.[@KTR] (generalized to $d$ dimensions).",
"\n\nEqs.(\\[avt\\]) and (\\[rghsc\\]) give the $v\\rightarrow 0$ scaling behavior of avalanche durations and interface roughness respectively for constant velocity driving. ",
"We note once again that these are for $\\xi>> L,$ [[*i.e.*]{} ]{}$v\\rightarrow 0$ before $L\\rightarrow\n\\infty.$ As mentioned before, even in critical phenomena for equilibrium phase transitions, as one approaches the transition for a fixed system size, one sees a change in the scaling form of dynamical variables. ",
"However, it is generally possible to obtain the behavior in this regime by requiring that there should be no dependence on (say) the reduced temperature $t$ for the behavior of a finite size system. ",
"This requirement comes from the fact that there are [*no*]{} thermodynamic singularities for a finite sized system: it is possible to go smoothly from one side of the phase transition to another. ",
"No such requirement exists for dynamical phase transitions, and one must be careful about possible $v$-dependence even in the $v\\rightarrow 0$ regime[@CDW].",
"\n\nThere are other mechanisms as well that can make the connection between RG exponents and apparent scaling difficult. ",
"For instance, for CDWs below the depinning threshold, the periodicity of the dynamical variable (the CDW phase) prevents it from advancing by more than $2\\pi$ anywhere in a single avalanche, but the same periodicity makes a region that has just avalanched susceptible to an imminent ‘retriggering’ of a fresh avalanche [@Middle]. ",
"The low frequency dynamics thus sees a non-trivial analog of the roughness exponent, and the distribution of avalanche sizes is singular as $v\\rightarrow 0.$ Another example is for interface roughness itself, where the steady state roughness over a subsystem of size $x$ in a system with size $L$ scales as $ w^2(x; L)\\sim \nx^2 L^{2\\zeta - 2}$ when $\\zeta > 1$ [@LL]. ",
"This is because the Fourier transform of the steady state roughness must be well behaved as $L\\rightarrow \\infty,$ and for $\\zeta > 1$ $w^2(x; L)$ is dominated by long wavelength modes with $q\\sim 1/L.$\n\nIn this paper we have shown that the renormalization group flows for depinning transitions are the same whether the system is driven with a constant force or a constant velocity. ",
"However, in the critical regime, the apparent scaling behavior of physical quantities can be different for the two driving modes. ",
"This is because scaling functions can depend on the system velocity in a singular manner as the transition is approached. ",
"This is unlike what is seen in equilibrium critical phenomena, and seems to be more common in the constant velocity driving mode. ",
"Despite this caveat, it is possible to obtain results for one driving mode from the other.",
"\n\nI thank Anne Tanguy and Maya Paczuski for useful discussions.",
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"\n\nIn Ref.[@KTR], using the model of Ref.[@TGR], the interface is in fact started with a flat configuration, but the same results are obtained starting from steady state (A. Tanguy, private communication). ",
"For the model of Ref.[@Sneppen], these results are [*only*]{} obtained starting from steady state [@SnepJen]. ",
"This is probably because the model of Ref.",
" [@Sneppen] is a ‘hybrid’ model, where only a single site moves at a time step, but can then trigger a brief avalanche (at the same time step). ",
"This is neither constant velocity nor constant force driving. ",
"In steady state, the avalanches have a characteristic size, and the model effectively has constant velocity driving. ",
"However, when one starts from a flat interface, the mean avalanche size rises from unity to its saturated value; in fact, all scaling in the short time regime for this case is quite imperfect. (",
"The connection between the changing mean avalanche size and the peculiar short time behavior of the roughness is seen better in the original version of the model [@Sneppen], where the initial pinning strengths are Gaussian distributed, but the new pinning strengths when sites move are uniform over $[0, 1)$.) Also, concepts of correlation lengths and compact avalanches are more tenuous [@LesTang] in this model.",
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"\n\nAvalanches can actually not be clearly separated in this regime, but it may be possible for continuum models to do this approximately by separating out fast and slow motion. ",
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[
"How To Create SSH Alias In Linux\n\nIf you frequently access a lot of different remote systems via SSH, this trick will save you some time. ",
"You can create SSH alias to frequently-accessed systems via SSH. ",
"This way you need not to remember all the different usernames, hostnames, ssh port numbers and IP addresses etc. ",
"Additionally, It avoids the need to repetitively type the same username/hostname, ip address, port no whenever you SSH into a Linux server(s).",
"\n\nCreate SSH Alias In Linux\n\nBefore I know this trick, usually, I connect to a remote system over SSH using anyone of the following ways.",
"\n\nI believe most of the newbie Linux users and/or admins would SSH into a remote system this way. ",
"However, If you SSH into multiple different systems, remembering all hostnames/ip addresses, usernames is bit difficult unless you write them down in a paper or save them in a text file. ",
"No worries! ",
"This can be easily solved by creating an alias(or shortcut) for SSH connections.",
"\n\nWe can create an alias for SSH commands in two methods.",
"\n\nMethod 1 – Using SSH Config File\n\nThis is my preferred way of creating aliases.",
"\n\nWe can use SSH default configuration file to create SSH alias. ",
"To do so, edit ~/.ssh/config file (If this file doesn’t exist, just create one):\n\nAgain make sure you have replaced the host, hostname, port number and ip address with your own. ",
"Save the file and exit.",
"\n\nThen, apply the changes using command:\n\n$ source ~/.bashrc\n\nOr,\n\n$ source ~/.bash_profile\n\nIn this method, you don’t even need to use “ssh alias-name” command. ",
"Instead, just use alias name only like below.",
"\n\n$ webserver\n$ dns\n$ dhcp\n$ ubuntu\n\nThese two methods are very simple, yet useful and much more convenient for those who often SSH into multiple different systems. ",
"Use any one of the aforementioned methods that suits for you to quickly access your remote Linux systems over SSH."
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[
"Questa, secondo Sky, è la situazione degli infortunati in casa rossonera in vista delle prossime partite:\n\nDONNARUMMA: Gigio ha ancora dolore, tra stasera e domani effettuerà degli esami per capire l'entità dello stiramento alla coscia destra. ",
"Salterà sicuramente il match di sabato contro la Juventus, ma ci proverà la successiva sfida contro la Lazio, anche se non sarà semplice.",
"\n\nPAQUETA': il brasiliano ha riportato una forte distorsione caviglia e già oggi pomeriggio farà dei controlli per capire l'entità del suo infortunio. ",
"La sensazione è che comunque dovrà restare fuori per qualche settimana.",
"\n\nCONTI: anche il terzino salterà la Juventus e come Donnarumma proverà a recuperare per il successivo impegno di campionato contro la Lazio.",
"\n\nKESSIE: l'ivoriano ha svolto oggi un lavoro personalizzato a Milanello e proverà a recuperare la trasferta di Torino.",
"\n\nSUSO: l'esterno spagnolo sta meglio, oggi si è allenato regolarmente a Milanello e quindi sarà a disposizione di Gattuso per il match contro la Juventus."
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[
"About\n\nWhat Lies in the Future?",
"\n\nWhat does the future hold for those who innovate? ",
"The road does not look smooth.",
"\n\nHistorically, America’s companies and its entrepreneurs depended on a reliable patent system to protect inventions. ",
"Today, despite a stream of technological innovation, our patent system has tipped away from protecting inventions to diminishing their value.",
"\n\nWith the implementation of AIA, the Patent Trial and Appeal Board has become a venue more focused on invalidating patents than encouraging innovation. ",
"Well-capitalized companies can infringe at will, waving the cudgel of proceedings at the PTAB at patentees who sue in federal district court for infringement. ",
"Supreme Court rulings in Alice and Mayo have made it harder for patentees to survive litigation, and both the district courts’ and the Federal Circuit’s implementation of those cases has been a study in docket control. ",
"Larger technology companies continue to fund media campaigns that paint smaller patent-holders as trolls unworthy of the title of patentee, a characterization that’s neither accurate nor fair.",
"\n\nIn short, holding a patent doesn’t mean much any more, unless you have many millions to defend it. ",
"Few do. ",
"How can innovators thrive in such an environment? ",
"What effects will these factors together have on American businesses, which depend on creativity and ingenuity to propel them to the front of global competition?",
"\n\nHopefully, by fighting the good fight for the right type of innovators, we can correct judicial misinterpretation of patents as well as amend the deleterious actions of Congress, and restore the historical value of patents for innovators. ",
"We need to protect innovation and preserve entrepreneurial businesses—large and small.",
"\n\nThat’s why I’m writing BusinessDeNovo: to comment on the changing tides of the rules around innovation and how they affect business. ",
"Fighting for patent-holders. ",
"Fighting for American innovation."
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"Toward scholarliness in doctoral dissertations: an analytical model.",
"\nThe dissertation is one of the most important vehicles for providing the novice researcher with the necessary socialization for a future of productive and scholarly research activities. ",
"Research productivity and the scholarliness of research are needed in developing nursing science. ",
"In this article, the processes that have the potential to enhance the quality of the doctoral dissertation are addressed; the psychological, social, and structural processes involved in completing a quality dissertation are delineated and discussed; and a model for the development of scholarliness and scholarship in doctoral programs is presented. ",
"The authors address three major areas for consideration: (a) the dependence of scholarship and scholarliness on researchable questions that are important, evolve in the context of theoretical considerations, and improve the scientific base of nursing knowledge; (b) the development of these qualities as enhanced by the resolution of several dilemmas that revolve around decisions made throughout the dissertation process; and (c) the recomendation that these qualities (scholarship and scholarliness) be fostered meticulously during doctoral education in nursing and, preferably, earlier."
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"Joseph Spector\n\nUSA TODAY Network-New York\n\nALBANY, N.Y. — New York will restrict sex offenders from using Pokémon Go and similar games, saying there are concerns the popular games could leave children vulnerable to predators.",
"\n\nNew York Gov. Andrew Cuomo on Monday directed the state Department of Corrections and Community Supervision to try to limit sex offenders from playing Pokémon Go.",
"\n\nHe also sent a letter to the game's software developer Niantic Inc. asking it to find ways to prohibit sex offenders from playing Pokémon Go.",
"\n\n“Protecting New York’s children is priority number one and, as technology evolves, we must ensure these advances don't become new avenues for dangerous predators to prey on new victims,\" Cuomo said in a statement.",
"\n\n'Pokémon Go' creators really want you to play safe\n\nOn Friday, New York state Sen. Jeff Klein, a Democrat from the Bronx, said he would introduce legislation to curb the use of the game by sexual predators.",
"\n\nThe concern is that the reality game can be used to lure players near the residences of sex offenders.",
"\n\nAlso, Klein said, a feature of the game can include a 'lure\" — where for a small fee a user can be directed to go to a specific location.",
"\n\nCuomo's order calls for the state corrections agency to install a new parole condition for sex offenders under state supervision to prohibit them \"downloading, accessing, or otherwise engaging in any Internet enabled gaming activities, including Pokémon Go.\"",
"\n\nN. J. man finds $2,000 cash playing Pokemon Go\n\nThe directive will impact about 3,000 Level 1, 2 and 3 sex offenders currently on parole, Cuomo's office said. ",
"According to the state Division of Criminal Justice Services, a Level 1 offender is someone with a \"low risk of repeat offense;\" a Level 2 sex offender has a \"moderate risk;\" and a Level 3 offender has a \"high risk of repeat offense and a threat to public safety exists.\"",
"\n\nThe state also will offer similar guidance to county probation offices, asking them to also adopt the policy.",
"\n\n\"Pokémon Go provided sex offenders with a virtual road map to our children,\" Klein said in a statement.",
"\n\n\"We know that pedophiles always seek new ways to lure victims and this new technology that entertains our kids, could also bring them close to dangerous individuals instead of Pokémon.\"",
"\n\nThe state Division of Criminal Justice Services also plans to work with Apple and Google on how to improve user safety.",
"\n\nJoseph Spector writes for Gannett's Albany (N.Y.) Bureau. ",
"Follow him on Twitter: @GannettAlbany\n\nThe bad and the ugly of Pokémon Go"
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"Science\n\nGive your little scientists the tools they need to make great discoveries. ",
"The curious mind will delight at Kaplan Toys' expansive offering of science activities. ",
"Build dinosaur bones, erupt a volcano or learn all about magnets. ",
"Kaplan Toys has everything the budding scientist needs to spark curiosity and wonder."
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"Evidence of three parallel evolutions of leaf dwarfism and phytogeography in Lysimachia sect. ",
"Nummularia in Japan and Taiwan.",
"\nPhylogenetic analyses were performed using nrITS variations of eight species including one variety of the genus Lysimachia sect. ",
"Nummularia in Japan and Taiwan, with another 34 Lysimachia species and Glaux maritime obtained from a DNA database. ",
"Among the four dwarf-leaf taxa in Japan and Taiwan, L. japonica var. ",
"minutissima of Japan and two Taiwanese species were placed into two different clades; and L. liukiuensis of Japan, was positioned outside of these two clades. ",
"These results suggest that three independent dwarfism events generated the four dwarf-leaf taxa; and that there were likely multiple migration events from China to Japan and Taiwan."
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[
"---\nabstract: 'We compute the influence of an external magnetic field on the Casimir energy of a massive charged scalar field confined between two parallel infinite plates. ",
"For this case the obtained result shows that the magnetic field inhibits the Casimir effect.'",
"\n---\n\n**[Bosonic Casimir effect in external magnetic field]{}**\n\nM.V. Cougo-Pinto[^1], C. Farina[^2], M. R. Negrão[^3] and A. C. Tort[^4]\n\n*Instituto de Física, Universidade Federal do Rio de Janeiro*\n\nCP 68528, Rio de Janeiro, RJ\n\n21945-970, Brazil\n\n1 true cm\n\nThe Casimir effect can be generally defined as the effect of a non-trivial space topology on the vacuum fluctuations of relativistic quantum fields [@MostepanenkoTrunovB; @PlunienMullerGreinerR]. ",
"The corresponding change in the vacuum fluctuations appears as a shift in the vacuum energy and an associated vacuum pressure. ",
"This shift is known as the Casimir energy of the field due to the given space constraints. ",
"The original Casimir effect [@Casimir48] is the attraction of two neutral perfectly conducting parallel plates placed in vacuum. ",
"The boundary conditions imposed by the metallic plates confine the vacuum fluctuations of the quantum electromagnetic field in the space between the plates. ",
"The effect of the boundary conditions can be viewed as a departure from the trivial topology of $\\R^3$ to the topology of $\\R^2\\times[0,a]$, where $a$ is the distance between the plates. ",
"The resulting shift in the vacuum energy of the quantum electromagnetic field was computed by Casimir and is given by[@Casimir48]: $$\\label{photonE}\n{\\cal E}_{\\gamma}(a)=-\\ell^2{\\pi^2\\over 720a^3}\\; ,$$ where $\\ell^2$ is the area of each plate and the close spacing between them is implemented by the condition $a\\ll\\ell$. The pressure corresponding to (\\[photonE\\]) was first measured by Sparnaay in 1958[@Sparnaay58] and more recently with high accuracy by Lamoreaux[@Lamoreaux97] and by Mohideen and Roy[@MohideenRoy98].",
"\n\nThe Casimir energy has become an important ingredient of any theory with non trivial vacuum and has been computed for fields other than the electromagnetic one with several types of boundary condition [@MostepanenkoTrunovB; @PlunienMullerGreinerR]. ",
"In the case of an electrically charged quantum field it poses by itself the question of how the charged fluctuations, and therefore the Casimir effect, are affected by fields coupling to the fluctuations through this charge. ",
"This question is strongly motivated by the fact that in a more complete picture of the Casimir effect the charged fluctuations of the constrained vacuum are or may be put under the influence of other fields. ",
"Within a hadron, for example, the vacuum fluctuations of quark fields are affected by the electromagnetic field of the quarks and by the color field of gluons and quarks. ",
"Also the vacuum fluctuations of gluon fields are affected by the color field of quarks and gluons. ",
"In this example of quarks and gluons the unavoidable influence of the fields on the constrained charged fluctuations is of extreme complexity. ",
"As a first step to the understanding of how charged fermionic and bosonic constrained vacuum fluctuations are affected by fields coupling to this charge we may consider the problem in its most simple form, to wit: vacuum fluctuations of Dirac or scalar electrically charged fields under the influence of an external constant uniform magnetic field and constrained by the simplest of the possible boundary conditions. ",
"In the case of a constrained Dirac vacuum the external field enhances the Casimir energy[@C-PFT98]. ",
"Here we shall consider a complex scalar field confined between two infinite plates with a constant uniform magnetic field in a direction perpendicular to the plates. ",
"The charged scalar field allows us to ignore kinematical complexities which are not relevant to an initial approach to this problem. ",
"The choice of a pure magnetic field excludes the possibility of pair creation for any field strength. ",
"The confinement between infinite plates is described by a simple form of Dirichlet boundary condition and the direction of the magnetic field perpendicular to the plates is obviously a simplifying choice. ",
"Under such assumptions the formalism may be kept simple in order for us to concentrate on the fundamental issue, which is the physical effect of the external field on the Casimir effect. ",
"Once the main feature of such influence is determined the path is open to consider more complicated geometries and external fields as well as other quantum vacua. ",
"Notice that we consider a problem in which the charged quantum vacuum is constrained by the boundary conditions and the external electromagnetic field is not. ",
"Therefore the influence of the external field on the charged vacuum already appears at the one-loop level, at which our calculations will be done. ",
"In contrast we have in the Scharnhorst effect [@Scharnhorst90; @Barton90; @BartonScharnhorst93; @C-PFST98a; @C-PFST98b] that boundary conditions on a pair of parallel plates are imposed on the electromagnetic quantum vacuum but not on the charged vacuum of electrons and positrons. ",
"As a result there is a change in the velocity of propagation of an external electromagnetic wave in the region between the plates. ",
"The Scharnhorst effect involves two-loop diagrams because the coupling between the external field and the quantum electromagnetic field requires the intermediation of a charged fermion loop.",
"\n\nLet us calculate the Casimir energy of the charged scalar field in a constant aplied magnetic field using a method introduced by Schwinger to obtain the Casimir energy [@Schwinger92] from the proper-time representation of the effective action[@Schwinger51]. ",
"Since the method has been clearly explained by Schwinger [@Schwinger92] and already applied to several situations [@C-PFSST] we may use it here without going into too much detail. ",
"We start with Schwinger’s proper-time formula for the effective action [@Schwinger51]: $$\\begin{aligned}\n{\\cal W} = - \\frac{i}{2} \\int_{s_0}^{\\infty}\n\\frac{ds}{s} Tr e^{-isH},\n\\label{acao}\\end{aligned}$$ where $s_{0}$ is a cutoff in the proper-time $s$, $Tr$ means the total trace and $H$ is the proper-time Hamiltonian, which is given by $(p -\neA)^{2} + m^{2}$, where $p_{\\mu} = - i {\\partial}_{\\mu}$, $e$ is the charge of the scalar field, $A$ is the electromagnetic potential and $m$ is the mass of the scalar field. ",
"The boundary condition gives for the component of the momentum which is perpendicular to the plates the eigenvalues $n\\pi/a$,where $n$ is a positive integer. ",
"The other spatial components of the momentum are constrained into the Landau levels created by the magnetic field ${\\bf B}$ and we choose the direction of ${\\bf B}$ in such a way that $eB$ is positive. ",
"The trace in (\\[acao\\]) is given by: $$\\begin{aligned}\nTr e^{-isH} = 2 e^{-ism^{2}}\\sum_{n'=0}^\\infty\n\\frac{{\\ell}^{2} eB}{2\\pi}\ne^{-iseB(2n'+1)} \\sum_{n=1}^\\infty e^{-is(n\\pi\n/a)^{2}} \\int \\frac{dtd\\omega}{2\\pi}\ne^{is\\omega^{2}}\n\\label{traco1}\\end{aligned}$$ where the factor $2$ is due to the two degrees of freedon in the complex field; the first sum is over the Landau levels with the corresponding multiplicity factor due to degeneracy; the second sum is over the eigenvalues stemming from the Dirichlet boundary conditions and the integral range is given by the measurement time $T$ and by the continuum of eigenvalues $\\omega$ of the operator $p^{0}$. Following Schwinger’s regularization prescription [@Schwinger92] we apply Poisson sum formula [@Poisson1823] to the second sum in order to obtain: $$\\begin{aligned}\n\\sum_{n=1}^\\infty e^{-is(n\\pi/a)^{2}} =\n\\frac{a}{\\sqrt{i\\pi s}}\n\\sum_{n=1}^\\infty e^{i(an)^{2}/s} +\n\\frac{a}{2\\sqrt{i\\pi s}} - \\frac{1}{2}.",
"\n\\label{poisum}\\end{aligned}$$ The sum over the Landau levels is straightforward and leads to: $$\\begin{aligned}\n\\sum_{n'=0}^\\infty \\frac{eB{\\ell}^{2}}{2\\pi}\ne^{-iseB(2n'+1)} = \\frac{eB{\\ell}^{2}}{4\\pi}\ncosech(iseB).",
"\n\\label{somaland}\\end{aligned}$$ Using (\\[poisum\\]) and (\\[somaland\\]) into (\\[traco1\\]), we obtain for the trace: $$\\begin{aligned}\nTr e^{-isH} = \\frac{a{\\ell}^{2}T}{4{\\pi}^{2}}\\frac{e^{-ism^{2}}}{is^{2}}[1 +\niseB{\\cal M}(iseB)] \\biggl [ \\frac{1}{2} - \\frac{\\sqrt{i\\pi s}}{2a} +\n\\sum_{n=1}^\\infty e^{i(an)^{2}/s} \\biggl ],\n\\label{traco2}\\end{aligned}$$ where ${\\cal M}$ is the function defined by: $$\\begin{aligned}\n{\\cal M}(\\xi) = cosech{\\xi} - {\\xi}^{-1}.",
"\n\\label{myfunc}\\end{aligned}$$ Substituting now equation (\\[traco2\\]) into equation (\\[acao\\]) we get the effective action: $$\\begin{aligned}\n{\\cal W} = {\\cal L}^{(1)}(B)Ta{\\ell}^{2} -\n{\\cal E}(a,B)T,\n\\label{acao2}\\end{aligned}$$ where on the right hand side the first term gives the (unrenormalized) effective Lagrangian: $$\\begin{aligned}\n{\\cal L}^{(1)}(B) = -\\frac{1}{16{\\pi}^{2}}\n{\\int_{s_{0}}}^{\\infty}\n\\frac{ds}{s^{3}} e^{-ism^{2}} (iseB)\ncosech(iseB)\n\\label{lag}\\end{aligned}$$ and the second term gives the (still cutoff-dependent) Casimir energy: $$\\begin{aligned}\n{\\cal E}(a,B) =\n-\\frac{a{\\ell}^{2}}{8{\\pi}^{2}} \\sum_{n=1}^\\infty\n{\\int_{s_{0}}}^{\\infty}\n\\frac{ds}{s^{3}} e^{-ism^{2} + i(an)^{2}/s} [ 1+ iseB{\\cal\nM}(iseB)],\n\\label{encas1}\\end{aligned}$$ which is the quantity we are interested in. ",
"The effective Lagrangian ${\\cal L}^{(1)}(B)$ is analogous to the Euler-Heisenberg Lagrangian for the fermionic case [@H-E] and was first obtained by Schwinger in 1951 [@Schwinger51]. ",
"Since it does not depend on $a$ it makes no contribution to the Casimir energy. ",
"Usually, spurious terms must be subtracted before eliminating the cutoff $s_o$ in (\\[encas1\\]) but in the present calculation they were all left in the effective Lagrangian which is of no concern to us here. ",
"So we may simply take $s_o=0$ in (\\[encas1\\]). ",
"Continuing with Schwinger’s method we now use Cauchy theorem to make a $\\pi/2$ clockwise rotation of the integration $s$-axis, which results in a substitution of $s$ by $-is$ in the integrand of (\\[encas1\\]). ",
"Part of this integrand can be expressed in terms of the modified Bessel function $K_{2}$ (cf. ",
"formula [**3471.9**]{} in [@GradshteijnRyzhik65]) and (\\[encas1\\]) reduces to: $$\\begin{aligned}\n\\frac{{\\cal E}(a,B)}{{\\ell}^{2}} &=& -\n\\frac{(am)^{2}}{4{\\pi}^{2}a^{3}} \\sum_{n\\in N}\\frac{1}{n^{2}}\nK_{2}(2amn) + \\nonumber\\\\\n&-& \\frac{1}{8{\\pi}^{2}a^{3}} \\sum_{n=1}^\\infty\n{\\int_{0}}^{\\infty} \\frac{ds}{s^3} e^{-s(am)^{2}-n^{2}/s} \ns\\,eBa^2{\\cal M}(s\\,eBa^{2}).",
"\n\\label{encas2}\\end{aligned}$$ The first term on the right hand side of this equation is the usual Casimir energy in the absence of the external magnetic field: $$\\begin{aligned}\n\\frac{{\\cal E}(a,0)}{a{\\ell}^{2}} = - \\frac{(am)^{2}}{4{\\pi}^{2}a^{4}}\n\\sum_{n=0}^\\infty \\frac{1}{n^{2}} K_{2}(2amn),\n\\label{encas0}\\end{aligned}$$ which is a result already known in current literature[@MostepanenkoTrunovB; @PlunienMullerGreinerR]; in the limit $m\\rightarrow 0$ this result reduces to (\\[photonE\\]) (actually because the complex scalar field has two degrees of freedon and the photon field two polarizations). ",
"Here we are interested in the second term on the right hand side of equation (\\[encas2\\]): $$\\begin{aligned}\n\\frac{{\\Delta\\cal E}(a,B)}{{\\ell}^{2}} =\n- \\frac{1}{8{\\pi}^{2}a^{3}} \\sum_{n=1}^\\infty\n{\\int_{0}}^{\\infty} \\frac{ds}{s^3} e^{-s(am)^{2}-n^{2}/s} \ns\\,eBa^2{\\cal M}(s\\,eBa^{2}).",
"\n\\label{Delta}\\end{aligned}$$ which measures the influence of the external magnetic field in the Casimir energy. ",
"Due to the simple behaviour of the function ${\\cal M}$ defined in (\\[myfunc\\]) we can determine the main features of this influence. ",
"The function $-\\xi\\,{\\cal M}(\\xi)$ increases monotonically from zero to the asymptotic value $1$ when $\\xi$ goes from $0$ to $\\infty$. Therefore we see that the external magnetic field always inhibits the Casimir energy of the scalar field and suppress it completely in the limit $B\\rightarrow\\infty$. This is the result that answer the question raised above. ",
"This result should be contrasted with the result for a Dirac field for which the Casimir energy is always enhanced by the external magnetic field [@C-PFT98]. ",
"It is very interesting that fermionic and bosonic charged vacuum present such a clear and opposite behaviours in presence of an external magnetic field. ",
"We didn’t find an intuitive explanation for those behaviours but it is quite possible that it is related with the paramagnetic and diamagnetic characters of fermionic and bosonic vacua, respectively. ",
"At any rate it is important to take in consideration this opposite behaviour of bosonic and fermionic vacua in the presence of an external field, because these vacua actually exist toghether in the presence of fields and may also be constrained by boundary conditions, as remarked above. ",
"Let us notice, for example, that the shift in the zero point energy caused by the external field depends on the mass of the field in the bosonic case (\\[Delta\\]) as well as in the fermionic case [@C-PFT98], and therefore a cancellation of zero point energies of those vacua depends on the specific relations between the masses of the quantum fields.",
"\n\nIt is also instructive to define $$\\label{m_B}\nm_B=\\sqrt{m^2+eB}$$ and write the complete Casimir energy (\\[encas2\\]) as: $$\\begin{aligned}\n\\frac{{\\cal E}(a,B)}{{\\ell}^{2}} =\n- \\frac{1}{8{\\pi}^{2}a^{3}} \\sum_{n=1}^\\infty\n{\\int_{0}}^{\\infty} ds\\,s^{-3} e^{-s(am_B)^{2}-n^{2}/s} \n\\frac{2s\\,eBa^2}{1-e^{-2s\\,eBa^2}}\n\\label{E(m_B)}\\end{aligned}$$ Comparing this expression with its limit when $B\\rightarrow 0$ we may say that the effect of the external magnetic field on the usual Casimir energy is given in the integrand of (\\[E(m\\_B)\\]) by the $B$ dependent fraction and the constant $m_B$ which appears in the exponential. ",
"When $B\\rightarrow 0$ the fraction tends to 1 and $m_B\\rightarrow m$. For a strong magnetic field the exponential is the dominant factor in the integrand and the effect of the magnetic field on the Casimir energy appears roughly as the substitution of $m$ by $m_B$; certainly the $B$ dependent fraction may still in this case affect the precise influence of the magnetic field on the Casimir energy.",
"\n\nLet us consider the strong field regime, in which changes in the charged vacuum should be more prominent. ",
"The integral in equation (\\[encas2\\]) is dominated by the exponential function whose maximum is $e^{-2amn}$ and occurs at $\\sigma = am/n$. Due to this feature we are justified in substituting the function ${\\cal M}(\\xi)$ by $2e^{-\\xi} - {\\xi}^{-1}$ if $B>>({\\phi}_{0}/a^{2})(a/{\\lambda}_{c})$, where ${\\phi}_{0}$ is the fundamental flux $1/e$ and ${\\lambda}_{c}$ is the Compton wavelength $1/m$. Therefore, in the strong field regime, the second term in (\\[encas2\\]) can also be expressed in terms of a modified Bessel function (formula [**3471,9**]{} in ref. [",
"@GradshteijnRyzhik65]), and the Casimir energy can be written as: $$\\begin{aligned}\n\\frac{{\\cal E}(a,B)}{{\\ell}^{2}} = - \\frac{eBa^2}{2{\\pi}^{2}a^{3}} \\sqrt{(am)^{2}\n+ eBa^{2}} \\sum_{n\\in N} \\frac{1}{n} K_{1}(2n\\sqrt{(am)^{2} +\neBa^{2}}).",
"\n\\label{encasf}\\end{aligned}$$ Notice that the sign in the square root is to be expected because in the regime we are working with a minus sign means energy creation or anihilation which cannot happen when we are dealing with a constant and uniform magnetic field. ",
"We can also use (\\[m\\_B\\]) to rewrite (\\[encasf\\]) in the following form: $$\\begin{aligned}\n\\frac{{\\cal E}(a,B)}{{\\ell}^{2}} = - eBa^2\\frac{am_B}{2{\\pi}^{2}a^{3}}\n\\sum_{n=1}^\\infty \\frac{1}{n} K_{1}(2am_B n)\\; ,\n\\label{encasf(m_B)}\\end{aligned}$$ which is in a more appropriated form to compare with (\\[encas0\\]). ",
"By further stressing the strong field regime we can take the asymptotic limit of $K_1$ (cf. [**",
"8446**]{} in [@GradshteijnRyzhik65]) in (\\[encasf(m\\_B)\\]) with $m_B\\approx eB$ to obtain: $$\\frac{{\\cal E}(a,B)}{{\\ell}^{2}} = \\frac{(eBa^2)^{5/4}}{a^3}e^{-2\\sqrt{eBa^2}}\\, .$$ Turning now our attention to the weak field regime $B<<({\\phi}_{0}/a^{2})(a/{\\lambda}_{c})$ we can substitute in the integrand of (\\[Delta\\]) $\\xi{\\cal M}(\\xi)$ by $-\\xi^2/6$ to obtain: $$\\frac{\\Delta{\\cal E}(a,B)}{{\\ell}^{2}} = -\\frac{(eBa^2)^2}{24{\\pi}^{2}a^{3}}\n\\sum_{n=1}^\\infty K_{0}(2am n)\\; ,\n\\label{encasfr}$$ Summarizing the results we have in equation (\\[encas2\\]) the exact expression for the influence of the external magnetic field on the Casimir energy of a scalar charged field. ",
"Equations (\\[encasf\\]) and (\\[encasfr\\]) particularize the result of equation (\\[encas2\\]) to the regimes of strong and weak magnetic field, respectively. ",
"In any case the external field inhibits the Casimir energy of the scalar field. ",
"This is in opposition with the case of a Dirac field, whose Casimir energy is enhanced by the external magnetic field [@C-PFT98]. ",
"We may also look at the interplay between constraints and external field on the quantum vacuum from a completely different point of view. ",
"Instead of asking what is the influence of the magnetic field on the Casimir energy of the constrained vacuum we can ask what is the effect that constraints on the vacuum have on the effective Lagrangian for the magneticf field. ",
"This study has already been done for the fermionic vacuum [@C-PFRT98] and will in the near future be presented also for the bosonic vacuum. ",
"It would also be interesting to investigate the effect of an external magnetic field on the bosonic vacuum of a scalar field with space-time symmetry given by the $\\kappa$-deformed Poincaré algebra [@C-PF97] in order to see the relation between the inhibiting effect of the magnetic field on the Casimir energy and the mechanism of creation of field excitations due to the deformation.",
"\n\n[\\*]{} Mostepanenko V M and Trunov N N 1997 [*The Casimir Effect and its Applications*]{} (Oxford: Clarendon)\\\nMostepanenko V M and Trunov N N 1988 [*Sov. ",
"Phys. ",
"Usp.*]{} [**",
"31**]{} 965 Plunien G, Muller B and Greiner W 1986 [*Phys. ",
"Rep.*]{} [**134**]{} 89 Casimir H B G 1948 [*Proc. ",
"Kon. ",
"Nederl. ",
"Akad. ",
"Wetensch.*]{} [**",
"51**]{} 793 Sparnaay M J 1958 [*Physica*]{} [**24**]{} 751 Lamoreaux S K 1997 [*Phys. ",
"Rev. Lett.*]{} [**",
"78**]{} 5 Mohideen U and Roy A 1998 [*Phys. ",
"Rev. Lett.*]{} [**",
"81**]{} 4549 Cougo-Pinto M V, Farina C and Tort A C 1998: [*Fermionic Casimir effect in external magnetic field*]{} (hep-th/9809215) Scharnhorst K 1990 [*Phys. ",
"Lett.*]{} [**",
"B236**]{} 354 Barton G 1990 [*Phys. ",
"Lett.*]{} [**",
"B237**]{} 559 Barton G and Sharnhorst K 1993 [*J. Phys.*]{} [**",
"A26**]{} 2037 Cougo-Pinto M V, Farina C, Santos F C and Tort A 1998 [*The speed of light in confined QED vacuum: Faster or slower than c?*]{} ",
"to be published in [Phys. ",
"Lett.]{} [**",
"B**]{} Cougo-Pinto M V, Farina C, Santos F C and Tort A 1998 [*QED vacuum between an unusual pair of plates*]{} (hep-th9811062) Schwinger J 1992 [*Lett. ",
"Math. ",
"Phys.*]{} [**",
"24**]{} 59 Schwinger J 1951 [*Phys. ",
"Rev.*]{} [**82**]{} 664 Cougo-Pinto M V, Farina C and Ségui-Santonja A J 1994 [*Lett. ",
"Math. ",
"Phys.*]{} [**",
"30**]{} 169 and [**31**]{} 309\\\nCougo-Pinto M V, Farina C and Tort A 1996 [*Lett. ",
"Math. ",
"Phys.*]{} [**",
"37**]{} 159 and [**38**]{} 337 Poisson S D 1823 [*Journal de l’Ecole Polytechnique*]{} [**XII**]{} (cahier XIX) 420 Heisenberg W 1935 [*Z. Phys.*]{} [**",
"90**]{} 209\\\nEuler H and Kockel B 1935 [*Naturwissensch.*]{} [**",
"23**]{} 246\\\nHeisenberg W and Euler H 1936 [*Z. Phys.*]{} [**",
"98**]{} 714\\\nWeisskopf V S 1936 [*K. Dan. ",
"Vidensk. ",
"Selsk. ",
"Mat. ",
"Fys. ",
"Medd.*]{}[**14**]{} 3 reprinted in Schwinger J 1958 [*Quantum Electrodynamics*]{} (New York: Dover) and translated in Miller A I 1994 [*Early Quantum Electrodynamics: a source book*]{} (Cambridge: University Press) Gradshteijn I S and Ryzhik I M 1965 [*Tables of Integrals, Series, and Products*]{} (New York: Academic Press) Cougo-Pinto M V, Farina C, Rafelski J and Tort A 1998 [*Phys. ",
"Lett.*]{} [**",
"B434**]{} 388 Cougo-Pinto M V and Farina C 1997 [*Phys. ",
"Lett.*]{} [**",
"B391**]{} 67\n\n[^1]: marcus@if.ufrj.br\n\n[^2]: farina@if.ufrj.br\n\n[^3]: guida@if.ufrj.br\n\n[^4]: tort@if.ufrj.br\n"
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"Wheelchair Accessible Vehicles\n\nWe are one of the main suppliers of wheelchair accessible contract hire agreements in the UK and have a team dedicated to creating competitive rentals at low payments.",
"\n\nThe specialist requirements are met by our expert consultants and quality vehicle converters. ",
"We boast:\n\na wide range of vehicles;\n\nlow initial payments;\n\nflexible rentals;\n\nfull maintenance options;\n\nhighest quality conversions.",
"\n\nWe are happy to chat through your requirements and show you how competitive we are in this specialist field. ",
"We have experience dealing directly with Care homes, Community Transport providers, Charities and local authorities. ",
"Long term relationships forged with the UK’s premier vehicle converters means we are in an ideal position to offer bespoke solutions to ensure vehicle specifications fully match our customers’ requirements.",
"\n\nWe offer a comprehensive maintenance package for every vehicle which covers the base vehicle and all items of the conversion including ramp or tail lift, steps, belts, tracking and auxiliary electrics. ",
"For vehicles fitted with tail-lifts we provide a six monthly service reminder and have national accounts with some of the UK’s leading tail lift service centres, ensuring the vehicle always meets the legal requirements of current legislation.",
"\n\nBy combining manufacturers’ discounts with competitively priced conversions we are able to offer packages to fit within required budgets. ",
"We are not tied to any particular converter or manufacturer meaning we can offer a flexible, informed solution to meet the needs of all transport providers.",
"\n\nIf you have any further questions on our Wheelchair Accessible Vehicle Contract Hire services contact us or call 0800 091 4252.",
"\n\nGet YourFree Fleet Guide\n\nWANT TO KNOW MORE?",
"\n\n0800 091 4252\n\nWHAT OUR CLIENTS SAY ABOUT US\n\n“We have worked with Toomey for the last eight years and during that time they have provided contract hire services for a growing proportion of our fleet. ",
"Despite having a relatively small fleet, through the high level of customer service provided by our account manager, Toomey have demonstrated that they really value our business in a way that larger providers have been unable to match.”."
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"Ausekla Zvaigzne (Star of Auseklis, Morning Star)\n\nAueskla means “star” in Lithuanian, and the Ausekla Zvaigzne (“star cross,” from the root Aust, dawn), or Star of Auseklis, is the emblem of the morning star in the Baltic region- the Latvian God Auseklis, as well as the Lithuanian goddess Ausrine, the daughter of the sun. ",
"The symbol is one of many ancient cosmological and magical symbols used in Eastern European folk art, and represents the god/goddess as the personification of the planet Venus.",
"\n\nIt probably should mean the same, yes. ",
"But as Auseklis is a Latvian symbol it has never been used as a sign on a house/ over the doorway. ",
"Jumis is the symbol our ancestors used on houses for protection and prosperity, not Auseklis. ",
"In this case you should look maybe for another type of cultural or mythological explanation.",
"\n\nActually you are wrong – it was used to protect house from evil spirits (in particular from Lietuvēns), it would be carved on or near doors or windows so the spirit cannot pass. ",
"Jumis, meanwhile, was used to decorate rooftops, which perhaps was much more obvious"
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"Project #3: Health Outcomes, Research Opportunities & the Direction of Research in Medicine One important reason for the dramatic increase in life expectancy over the past century is the development of new scientific and biomedical knowledge. ",
"Despite the importance of such research, there has not been any comprehensive attempt to measure the link between medical research and the subsequent improvements in health. ",
"There has also been little scholarly interest in the most productive allocation of limited research dollars across researchers. ",
"Young researchers may be better positioned to direct their research to areas with increasing health needs but recognizing areas with the most productive research opportunities may require the wisdom of experienced researchers. ",
"Uncovering these responses is important for predicting how aging of US biomedical researchers will alter research and optimal policy. ",
"In the first part of this project we estimate how biomedical research communities respond to shocks to research opportunities and population health needs. ",
"We will examine how different researcher characteristics such as career stage, availability of funding, and collaborators' characteristics influence the link between topic choice and disease prevalence as well as the link between topic choice and the quality of research opportunities. ",
"A weak response by researchers of a particular type would mean that the researchers set their own agenda in a way that does not take the social need into account. ",
"In the second part of this project we estimate the direct and indirect impacts that research effort on a disease has on health outcomes. ",
"We will compare changes in the extent of research effort across diseases with subsequent changes in disease-specific health outcomes. ",
"The analyses take into account health care expenditures, lifestyle and demographic changes, and for-profit medical innovation as factors that influence health outcomes. ",
"We will also estimate the indirect impacts that basic and more applied forms of medical research have on health outcomes through their impacts on for-profit medical innovation. ",
"The results will form an important input into calculating the optimal level of private and public funding of medical research. ",
"RELEVANCE (See instructions): Life expectancy has increased dramatically over the past century. ",
"This project uncovers the role of medical research in delivering these advances in life expectancy. ",
"The project also examines how population health needs influence the choice of research topics at different career stages, which will help predict how the aging of US biomedical researchers will impact research and its health benefits."
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"I installed maven manually i.e. unpacked, moved to /opt/maven.",
"\nMy /etc/environment file\nJAVA_HOME=\"/opt/java/jdk1.6.0_31\"\nJRE_HOME=\"/opt/java/jdk1.6.0_31/jre\"\nMAVEN_HOME=\"/opt/maven\"\nPATH=\"/usr/local/sbin:/usr/local/bin:/usr/bin:/sbin:/bin:/usr/games:$JAVA_HOME:$JRE_HOME:$MAVEN_HOME/bin\"\n\nWhen I replace in PATH $MAVEN_HOME with /opt/maven everything works. ",
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"It is read by the pam_env module during log in, and it doesn't do variable expansions, it only accepts KEY=VALUE pairs. ",
"You could instead edit /etc/profile (which is a shell script), and add\nPATH=$PATH:$JAVA_HOME:$JRE_HOME:$MAVEN_HOME/bin\n\n"
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"The relationship between arm-ankle pressure difference and peak systolic velocity in patients with stenotic lower extremity vein grafts.",
"\nThe relationship between the measured arm-ankle pressure difference (AAPD), or the ankle/arm index (AAI), and the focal peak systolic velocity (PSV) at stenotic sites of infrainguinal vein grafts has not been determined. ",
"We attempted to relate these two parameters. ",
"We used Doppler systolic pressures and duplex ultrasonography to study 35 infrainguinal vein bypass grafts followed in a surveillance protocol. ",
"The following graft groups were identified: grafts in nondiabetic patients (n = 26), grafts in diabetic patients (n = 9), nonrevised stenotic grafts (n = 14), revised stenotic grafts (n = 14), and normal grafts (n = 7). ",
"AAPD and AAI were measured in both lower extremities. ",
"Pressure gradients across graft stenoses were indirectly estimated using the modified Bernoulli equation (delta P =4V2). ",
"Measured AAPDs and estimated pressure gradients showed moderate correlation in nondiabetic (r = 0.58) and diabetic (r = 0.63) patients. ",
"Correlation was fair (r = 0.3) prior to graft revision. ",
"There was no correlation (r = 0.1) in the nonrevised stenotic grafts. ",
"For individual patients with stenotic grafts who were followed in consecutive visits, the correlation varied from none to good (r range 0.01 to 0.71). ",
"We conclude that there is a lack of consistent correlation between the measured AAPD, or AAI, and the estimated stenotic graft pressure gradient. ",
"This finding illustrates the limitation of the AAI as a monitoring test to predict failure of stenotic infrainguinal vein grafts."
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"\n\nHi,\nI am glad to work with you on your project.",
"I understood your requirements that you have added to the project description.",
"\nwhen you message me then we talk more the project.",
"I am confident to complete your projecLisää\n\nHello Sir\nI am very interested in creating Presentation for Engineering subject.",
"\nI have 5+ years experience and expertise in making professional presentations, pitch-deck presentations, product presentations, proLisää\n\nI am a PowerPoint and keynote presentation specialist. ",
"Basically i work outside market place so I don't have any re view here. ",
"But i have a vast experiences on this.",
"\nI believe in work so please take a look of my samplLisää\n\nI am a perfectionist, have a great passion for Arts and Loves new challenges. ",
"Have a great knowledge in design, planning, estimating, cost evaluation for civil and especially structural design.",
"\nRelevant Skills and ExpLisää"
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"inSANE\n\nGame Summary\n\nTHQ Inc. has entered into a multi-year agreement with Guillermo del Toro, writer and director of Hellboy and the Oscar-winning Pan's Labyrinth, to create inSANETM, an original trilogy of triple-A games. ",
"To be developed by Volition, Inc.®, inSANE will deliver an unprecedented cinematic experience while redefining storytelling in gaming. ",
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"Q:\n\nDoes the Matsya Purana forbid one to read Rajasika and Tamasika Puranas?",
"\n\nPadma Purana, Uttara Khanda, Chapter 236 states:\n\nसात्विका मोक्षदाः प्रोक्ताः राजसा सर्वदा अशुभाः ।",
"\nतथैव तामसा देवि निरयप्राप्तिहेतवः ।।",
"\nAmong these puranas Satvika Puranas lead one to liberation, Rajasa do not cause good and Tamasa puranas lead one to hell. (",
"236.2)\n\nNow, this guna based classification of Puranas, I have been told, is also present in the Matsya (which is a Tamasika Purana incidentally in the Padma Purana). ",
"I want to know whether the Matsya Purana does indeed mention a classification of the Puranas on the basis of gunas. ",
"If yes, does it discourage one to read Rajasika and Tamasika Puranas?",
"\n\nA:\n\nThe Matsya Purana indeed classifies the eighteen Puranas according to gunas but it doesn't explicitly discourage one to read Rajasika and Tamasika Puranas or even state them to be inferior to Sattvika Puranas:\n\nThe glory of Hari is greater in Sattvika Puranas; the glory of Brahma is greater in Rajasika Puranas; and that of Agni and Shiva greater in Tamasika Puranas. ",
"In mixed Puranas, the glory of Sarasvati and the pitrs is said to be greater. (",
"Matsya Purana, Chapter 53, Verses 67-68)\n\n"
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"ContainerNode, announceProgress bool) *Spec {\n\tspec := &Spec{\n\t\tsubject: subject,\n\t\tcontainers: containers,\n\t\tfocused: subject.",
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"Samples(),\n\t\tComponentTexts: componentTexts,\n\t\tComponentCodeLocations: componentCodeLocations,\n\t\tState: spec.getState(),\n\t\tRunTime: runTime,\n\t\tFailure: spec.failure,\n\t\tMeasurements: spec.measurementsReport(),\n\t\tSuiteID: suiteID,\n\t}\n}\n\nfunc (spec *Spec) ConcatenatedString() string {\n\ts := \"\"\n\tfor _, container := range spec.containers {\n\t\ts += container.",
"Text() + \" \"\n\t}\n\n\treturn s + spec.subject.",
"Text()\n}\n\nfunc (spec *Spec) Run(writer io.",
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"Since(spec.startTime)\n\t}()\n\n\tfor sample := 0; sample < spec.subject.",
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"SetupNodesOfType(types.",
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"= types.",
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"SpecStatePassed {\n\t\t\t\t\tspec.state = justAfterEachState\n\t\t\t\t\tspec.failure = justAfterEachFailure\n\t\t\t\t}\n\t\t\t}\n\t\t}\n\n\t\tfor i := innerMostContainerIndexToUnwind; i >= 0; i-- {\n\t\t\tcontainer := spec.containers[i]\n\t\t\tfor _, afterEach := range container.",
"SetupNodesOfType(types.",
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"SetupNodesOfType(types.",
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"BasicNode) {\n\tif spec.announceProgress {\n\t\ts := fmt.",
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"Failed() {\n\t\treturn map[string]*types.",
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"MeasureNode).MeasurementsReport()\n}\n"
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[
"1. ",
"Introduction {#sec1-insects-11-00256}\n===============\n\nPopulation growth rate can be affected by positive density dependence, which is also known as inverse density dependence or the Allee effect at low population density, whereby there is a positive relationship between population density and growth rate. ",
"\\[[@B1-insects-11-00256],[@B2-insects-11-00256],[@B3-insects-11-00256]\\]. ",
"Allee effects may cause longer lag times for population growth, and subsequently slow the growth and spread rate and lower the likelihood of persistence of both invasive populations and populations under conservation management \\[[@B4-insects-11-00256],[@B5-insects-11-00256]\\]. ",
"Although an Allee threshold suggests a definitive value, in reality, the threshold is probabilistic, with a higher probability of population growth above the threshold and probable reduction to extinction below. ",
"Moreover, the thresholds may vary depending on spatial and temporal ecosystem factors \\[[@B6-insects-11-00256]\\]. ",
"Allee effects may be due to several factors, including reduced mating, which may become problematic for low density populations \\[[@B7-insects-11-00256]\\]. ",
"Positive density dependence may be exploited for population management where not all of the population necessarily needs to be killed to achieve eradication \\[[@B8-insects-11-00256]\\], or to ensure that population density is sufficient for ongoing continuation of a population \\[[@B9-insects-11-00256],[@B10-insects-11-00256]\\].",
"\n\nTo date, Allee effects in insects have been quantified mostly from large-scale multi-year population studies. ",
"For example, reduced reproductive success and increased predation of low-density Monarch butterfly *Danaus plexippus* populations \\[[@B9-insects-11-00256],[@B11-insects-11-00256]\\]. ",
"Another study has estimated a trap-catch-based Allee threshold for gypsy moth *Lymantria dispar dispar* from multi-year trapping data from multiple trap transects placed through the invasion front of the moth in North America \\[[@B8-insects-11-00256],[@B12-insects-11-00256]\\]. ",
"Such trials use extensive field datasets and analyses to understand what factors promote or have the greatest effect on population persistence.",
"\n\nTrying to assess the effect of spatial separation on mate finding and mating success is difficult in the laboratory as space is limited. ",
"Study systems must be large enough to achieve sufficient distance between individuals, but small enough that the individuals can still be tracked by the observers. ",
"The advent of 3D printers has enabled the production of complex shapes, such as mazes, to be readily created. ",
"Mazes of known mathematical complexity can be readily used for hypothesis testing \\[[@B13-insects-11-00256]\\]. ",
"To test the concept, we investigated whether mazes of varying complexity could be used to increase spatial separation to investigate mate-finding Allee effects in the laboratory using *Drosophila simulans*.",
"\n\nMazes have often been used to investigate learning in organisms (e.g., heat mazes in *Drosophila melanogaster* \\[[@B14-insects-11-00256]\\] and water mazes for rats \\[[@B15-insects-11-00256]\\]). ",
"However, to the best of our knowledge, mazes have not been used to modify spatial distance between individuals. ",
"We hypothesised that mazes of increasing complexity would increase the time taken for one male and one female *Drosophila simulans* placed at opposite ends of the maze to find each other, artificially increasing the Allee threshold. ",
"This is due to an increase in the length of the route needed to be taken through the maze, reducing the probability of mating occurring within a fixed time. ",
"Furthermore, we hypothesised that the presence of a generalist pseudoscorpion predator (*Chelifer cancroides*) in the maze would increase the time taken for pairs of flies to find each other, again increasing the Allee threshold. ",
"However, we predicted that the presence of multiple pairs of flies in the maze, increasing fly density, would overcome the negative effects of the spatial separation and predation on mate finding and frequency of mating.",
"\n\n2. ",
"Materials and Methods {#sec2-insects-11-00256}\n========================\n\n*D. simulans* colonies were maintained on a diet of potato flakes (128 g), casein (28 g), castor sugar (20 g), baking yeast (8 g), methyl paraben (0.6 g), ascorbic acid (1.3 g) and benzoic acid (0.6 g) at 25 °C in ambient day length. ",
"Twenty millilitres of the diet/oviposition material was added to a 120 mL circular vial (110 mm × 43 mm i.d.), ",
"and 20 mL of water was added to hydrate the diet/oviposition material. ",
"As *D. simulans* pupate out of the diet on the edge of the vial, the vials were lined with waxed baking paper so that pupae could easily be removed. ",
"To facilitate pupal removal, the baking paper was sprayed lightly with water and each pupa was placed into a polycarbonate test tube (75 mm long, 10 mm i.d.) ",
"stoppered with a water-moistened cotton wick. ",
"Pupae-filled tubes were kept in a sealed bag to maintain high levels of humidity. ",
"No food was added to these tubes. ",
"Tubes were checked once daily for eclosion. ",
"On eclosion, the adults were sexed and 30% (by volume) sugar water was added to the cotton wick. ",
"Adult flies were used once when they were between 48 and 72 h old, when the majority of *Drosophila spp.* ",
"are sexually receptive \\[[@B16-insects-11-00256]\\].",
"\n\n2.1. ",
"Chelifer cancroides Rearing {#sec2dot1-insects-11-00256}\n--------------------------------\n\nThe *Chelifer cancroides* colony of many hundred individuals was maintained on a diet of *D. simulans* and cultured following Read et al. ",
"\\[[@B17-insects-11-00256]\\]. ",
"The *D. simulans* were maintained as above. ",
"When *D. simulans* started to pupate in the vials, an additional 20 mL of water was added and the larvae/diet solution was gently mixed so that it could be poured into the *C. cancroides* colony weekly. ",
"The *C. cancroides* were observed feeding on adult flies but may have also fed on the larvae or the diet itself. ",
"For the trials using *C. cancroides*, five sub-colonies of six males and six females were developed from the main colony; one colony for each day of the working week. ",
"Sub-colonies were fed a diet of \\~20 mixed sex adult *D. simulans* once a week immediately after participating in trials. ",
"The following day all dead and live flies were removed from the colony so that the colony was starved for six days prior to use. ",
"They had access to water ad libitum and tissue paper to hide in. ",
"The colonies were maintained at 25 ± 2 °C in ambient day length.",
"\n\n2.2. ",
"Maze Creation Overview {#sec2dot2-insects-11-00256}\n---------------------------\n\nTwo-dimensional mazes with variable complexity were generated on the website, [www.mazegenerator.se](www.mazegenerator.se). ",
"Circular mazes were set to have an \"outer diameter\" of 20 cells (90 mm). ",
"The \"internal diameter\" setting varied with the desired complexity. ",
"Four maze complexities were generated: simple, with an external and internal diameter of 20 cells (only an outer wall, similar to a Petri dish); easy, with an external diameter of 20 cells, internal diameter of 14 cells (three cells of maze); medium, with an external diameter of 20 cells, internal diameter of eight cells (six cells of maze); and hard, with an external diameter of 20 cells and an internal diameter of four cells (eight cells of maze). ",
"Adjacent walls (cells) were 3 mm apart and the height of the maze was 5 mm, and a Perspex lid was added to the top of the maze to prevent fly escape and facilitate observations. ",
"Two additional parameter values---elitism and river---were set to zero in the maze generator programme. ",
"The elitist value controls the solution length of the maze. ",
"The non-elitist (elitism = 0) parameter weights the maze generator programme to generate a maze that has a solution path that goes through a large portion of the maze, conversely an elitist maze (elitism = 100) has a short solution length from the edge to the centre of the maze. ",
"The river value controlled the length of dead-ends. ",
"A high river value (river = 100) weights the programme to generate a maze with few but long corridors that lead to a dead end, whereas a low river value of 0, as used here, has many short corridors that lead to dead ends ([Figure 1](#insects-11-00256-f001){ref-type=\"fig\"}). ",
"Technical details are provided in [Appendix A](#app1-insects-11-00256){ref-type=\"app\"}.",
"\n\n2.3. ",
"Spatial Complexity Trial {#sec2dot3-insects-11-00256}\n-----------------------------\n\nA single pair of flies was tested to determine whether the time to find a mate and the probability of mating changed with increasing spatial separation, caused by the escalating complexity of the maze. ",
"In these microcosms, flies were given one hour to find each other and mate. ",
"Males were released through the hole in the centre of the maze and females through the side entrance generated by the maze generator. ",
"The entrances were then closed with masking tape. ",
"The four complexity levels (simple, easy, medium and hard) were tested simultaneously and replicated 30 times. ",
"Time taken for mate location was recorded, as well as whether the pair mated. ",
"This was assessed visually and any pairs mating were recorded as successful if the pair remained in copula (minimum 14 and maximum 26 min in copula recorded), without the female trying to avoid the male.",
"\n\n2.4. ",
"Fly Density {#sec2dot4-insects-11-00256}\n----------------\n\nWe tested whether there was an increase in the probability of mating and a reduction in the time for pairs to find each other when there was a higher density of flies in the maze. ",
"One, three and five pairs of flies were tested in the \"easy\" solution maze. ",
"In addition, to determine whether there would be any effect of a predator on mate finding and mating, we added a 6-day starved male *Chelifer cancroides*. ",
"The predator was added to the halfway point between both the male and female maze entry points immediately prior to the addition of the flies. ",
"Each replicate comprised one of each of the three fly density levels with and without a predator (six mazes total per replicate). ",
"After the predator was added, male flies were placed in the centre of the maze and held there using a thin (1 mm) gate inserted through the Perspex lid until all female flies were added to the maze through the side entrance. ",
"When all flies were present in the maze, the males were allowed to leave the centre and the trial was started and ran for a maximum of one hour as above. ",
"All six treatments were run simultaneously for each replicate (n = 30 replicates). ",
"Time for the first pair to find each other was recorded, as well as any predation, sex of the predated fly and the number of pairs mating following the criteria above.",
"\n\nThe minimum distance that the flies had to travel to meet each other was investigated by using the tracking software MaxTRAQ v1.91 (Innovision Systems, Lapeer, MI, USA) ([Figure 2](#insects-11-00256-f002){ref-type=\"fig\"}). ",
"The distance to solve the maze from edge to centre was measured and the distance was divided by two assuming that both insects would move towards each other at the same rate.",
"\n\n2.5. ",
"Statistical Analysis {#sec2dot5-insects-11-00256}\n-------------------------\n\nTo model the effect of spatial complexity on maze solution length and percentage of single-pair fly trials where flies found each other, we performed a linear regression using Origin v. 8.5 (OriginLab Corporation, Northampton, MA, USA). ",
"To analyse the effect of the number of pairs of flies and the presence of a predator on time to find a mate, and the probability of mating occurring, we used a Generalized Linear Model (GLM) with a Poisson distribution logarithm-link function, and a GLM with a binomial distribution logit-link function, respectively. ",
"The GLM analyses were performed in GenStat v. 17 (VSN International Ltd., Hemel Hempstead, UK).",
"\n\n3. ",
"Results {#sec3-insects-11-00256}\n==========\n\nThe flies failed to perform in the way we anticipated as single pairs, with only three of the 30 pairs of flies mating in the simple maze and one pair of flies mating in the medium complexity maze. ",
"No flies mated in the easy or hard complexities in the one hour allotted. ",
"However, as we predicted, fly mating rates increased with increasing fly density.",
"\n\n3.1. ",
"Spatial Complexity {#sec3dot1-insects-11-00256}\n-----------------------\n\nThe maze experiments showed a positive linear relationship between the complexity of the maze and the solution length (R^2^ = 0.984). ",
"The minimum distance flies needed to travel to meet each other increased by 7-, 10- and 15-fold for flies in the easy, medium and hard mazes, respectively, compared to the flies in the simple maze.",
"\n\nThe maze complexity also affected the percentage of flies that found a mate in the 1 h period (R^2^ = 0.957) in the single pair treatments ([Figure 3](#insects-11-00256-f003){ref-type=\"fig\"}).",
"\n\nThe probability that a pair of flies would find each other was significantly different between the treatments as assessed by a Generalized Linear Model (GLM), binominal distribution logit-link (deviance = 12.6, 141.2; d.f. ",
"= 1; *p* \\< 0.001) for the main interaction for the complexity of the maze, explaining approximately 91% of deviance observed. ",
"Of the flies that found each other, the time taken to do so also increased with an increase in the complexity of the maze; GLM-Poisson log-link (deviance = 173, 1403; d.f. ",
"= 1; *p* \\< 0.001) main interaction of the complexity on time to find each other explaining 87% of the deviance. ",
"However, this difference was due to the difference between the simple maze and the three other complexities only; *p* = 0.115 for the comparison between the time taken in the easy, medium and hard complexities ([Figure 4](#insects-11-00256-f004){ref-type=\"fig\"}).",
"\n\n3.2. ",
"Fly Density {#sec3dot2-insects-11-00256}\n----------------\n\nThe average time for one fly of each sex to find the other varied with the number of pairs that were in the maze and whether there was a predator present ([Table 1](#insects-11-00256-t001){ref-type=\"table\"}). ",
"The time for initial mate finding was faster in the multiple pair fly treatments than for the single pair of flies. ",
"The presence of a predator in the maze increased the time it took for all flies to find each other ([Table 2](#insects-11-00256-t002){ref-type=\"table\"}), with a total of 18 male and 15 female flies killed by the predators during the experiment (<https://youtu.be/BEaRsO0h2v0>).",
"\n\nDespite increasing the time it took for individuals to find each other in the maze, the presence of a predator did not reduce the probability that at least one pair would mate. ",
"There was no effect of the interaction term of the predator presence and number of pairs mating, so this was removed from the analysis and main effects only are shown ([Table 3](#insects-11-00256-t003){ref-type=\"table\"}). ",
"The biggest effect on mating was the number of pairs present in the maze, with an increase in the percentage of flies mating as the density of flies in the maze increased.",
"\n\n4. ",
"Discussion {#sec4-insects-11-00256}\n=============\n\nMazes of increasing complexity increased the spatial distance between individual *D. simulans*, providing a baseline for testing mate-finding effects in this system. ",
"As complexity (solution length) increased, the initial distance between flies increased, requiring individual flies to spend a prolonged amount of their finite time searching for each other. ",
"By reducing encounter rates in the limited area of the 90 mm diameter arena via increased spatial separation, we expected to manipulate the mate-finding Allee threshold relative to the Petri dish style simple maze.",
"\n\nThe probability that flies would find each other within the one hour allowed for the flies to navigate the maze decreased with increasing complexity, with all flies finding each other when there was only an outer wall present (similar to a Petri dish). ",
"Over half of the flies did not find each other in the most complex maze offered; a maze that had a route that was 15-fold longer than the simple Petri dish style maze.",
"\n\nAt the individual fly level and replicate \"population\" level, the increase in fly density was able to overcome the spatial separation effects and the effect of the single predator in the system. ",
"The addition of multiple flies increased fly density in the mazes, the percentage of females mating and percentage of any mating event at all occurring per replicate in both the predator present and predator absent trials. ",
"In the single pair treatments, only 73% of trials had the potential to have a mating event occur because the predator killed flies in eight of the 30 trials. ",
"Even though a greater number of flies overall were killed in the multiple pair fly trials, flies were killed at a similar rate between the different fly density treatments, and all multiple pair fly trials had flies of each sex remaining at the end of the trial. ",
"We recognize that by printing mazes that had open corridors of approximately 3mm width with a height of 5 mm, there was limited ability for the Drosophila to use flight, a prime mode of their locomotion for predator avoidance. ",
"However, as we were investigating the role of habitat complexity and fly density on mate finding ability and subsequent copulation, their reliance on short hops and running here was considered sufficient for these treatments. ",
"The increase in the number of flies in the mazes increased the density of the flies, thus increased the probability of mating occurring, but it may have also promoted an increased rate of dispersal from the respective release area for the males and due to perceived competition \\[[@B18-insects-11-00256]\\]. ",
"We did not measure parameters such as walking speed or area of maze traversed to estimate dispersal rate to determine whether this phenomenon could have been a factor that influenced time to mate finding rates in our trials.",
"\n\nIn the single pair trials, the presence of the predator increased the time that was taken for the pair of flies to meet. ",
"This reduced the probability that mating would occur partly through direct predation and because of the increased searching time required in the presence of the predator potentially altering fly behaviour in order to avoid the predator \\[[@B19-insects-11-00256]\\]. ",
"When the predator was present only one of the single pair populations or 3% of the replicates had a mating event, whereas 43% and 46% of the three and five pair populations, respectively, were successful at mating. ",
"Under the trial conditions of only one hour to perform mating, the higher fly density populations would be more likely to persist than the single pair populations. ",
"This increase in fly density led to a greater chance of finding and successful mating by pairs and a reduced likelihood of attack potentially because of a Type II functional response with satiation of the natural enemy \\[[@B20-insects-11-00256]\\]. ",
"A difference in prey handling time or multiple predators would probably shift the fly density requirement up to maintain a population as more prey would probably be removed from the population potentially leading to a decline in the population growth rates \\[[@B21-insects-11-00256]\\]. ",
"Further, non-mortal sources of reduced mating can be caused through the perception of a predator via senses such as smell and vibration that induce avoidance responses in *Drosophila*, which can result in behaviours that depress the per capita population growth rate \\[[@B19-insects-11-00256]\\]. ",
"The presence of the predator used here, *C. cancroides*, in the maze did increase the time taken for the first pairs of flies to find each other, but not appear to have an effect on the probability of fly mating, unlike the increase in fly density that lead to 6 or 8 times the probability of any mating occurring in the three and five pair trials respectively, than was observed in the single pair trials.",
"\n\nAn increase in spatial separation between the sexes increased the time taken for individual flies of the opposite sex to find each other from simple to easy, but not for the easy--hard complexities. ",
"The initial change in spatial separation from the simple to easy maze was 13 cm distance or an increase ×6.9 the distance. ",
"Further, for the males to escape the centre of the maze they were required to find the 3 mm gap in the inner wall to enter the walled section of the 3D-printed maze. ",
"The increase in distance from easy to hard complexity was 18.1 cm or ×2.2 the distance. ",
"This distance may have been insufficient to observe noticeable changes in the time taken for the flies to navigate the maze. ",
"Further, once flies were in the walled section of the maze they may have quickly negotiated the corridors. ",
"Unfortunately, we did not quantify arrestment behaviours of the flies related to the sections of maze they were in.",
"\n\nKramer et al. ",
"\\[[@B22-insects-11-00256]\\] used a meta-analysis approach on data from natural populations and concluded that the most common sources of Allee effects were from mate limitation, reduction in cooperative defence, reduced predator satiation, cooperative feeding or over-dispersal and change in habitat, concurring with earlier authors \\[[@B1-insects-11-00256],[@B5-insects-11-00256],[@B23-insects-11-00256],[@B24-insects-11-00256]\\]. ",
"The maze used here offers a simple way to assess the fundamental effects of mate limitation and predator satiation on population growth. ",
"Mazes could be used for a large number of other behavioural applications including traditional aspects of organism learning. ",
"The 3D printed mazes can also work for larger organisms; a pilot study assessed a larger version of the maze for Queensland fruit fly, *Bactrocera tryoni*. ",
"Here, we have observed and tested the role of mate limitation through varying fly density and predator satiation or predator-induced avoidance behaviours exhibited by the flies in a novel use of 3D printed mazes of varying complexity.",
"\n\n5. ",
"Conclusions {#sec5-insects-11-00256}\n==============\n\nFlies were able to successfully navigate and mate during their natural dusk mating period. ",
"By overhead recording of the maze trials, any assessment of behavioural responses to non-visual stimuli can be assessed with image analysis. ",
"Natural dispersal was limited in the maze system, as the flies were forced to walk rather than fly. ",
"Our next step is to determine the correlation between maze complexity and real world distance, in order to model real-world spatial separation effects from laboratory-derived datasets.",
"\n\nThe authors thank Tom Sullivan for insect rearing and formulation of the diet, Ron van Toor for supplying the *Chelifer cancroides* colony and sharing his insect husbandry and behaviour knowledge and Donna Gibson and Martin Heffer for help with filming and editing the video sequence. ",
"The authors would also thank two anonymous reviewers for reviewing and improving this manuscript. ",
"AJP thanks Instituto Politécnico Nacional and CONACYT, México and the authors thank the Better Border Biosecurity (B3.net.nz) collaboration.",
"\n\nL.D.S., A.J.-P., J.R.B., J.M.K. and D.M.S. designed the concept and hypotheses. ",
"L.D.S., N.J.S., J.F. and R.W. collected the data and improved on the methodology. ",
"L.D.S. led the writing of the manuscript as part of a PhD project supervised by D.M.S., J.M.K. and J.R.B. All authors have read and agreed to the published version of the manuscript.",
"\n\nThis research received no external funding.",
"\n\nThe authors declare no conflicts of interest.",
"\n\n**Maze creation detail**\n\nTo make a 3D maze from the 2D maze generated by the website (mazegenerator.se), the 2D maze was exported as a scalable vector graphics file (.svg) (without a solution shown) from the maze generator website. ",
"This file was opened in Inkscape v.0.91 (Free Software Foundation Inc. Boston MA., ",
"USA). ",
"In Inkscape, the walls in the maze were initially broken into sections and were treated as separate entities or \"paths\". ",
"To create a single path (wall) for editing, the maze image was combined (ctrl-K). ",
"The edit paths by nodes option was selected to create an outline of the maze walls, allowing the user to visually confirm all walls were selected thus had a continuous outline. ",
"When the outline was confirmed, the stroke to path option was selected to create the solid walls of the maze. ",
"At this point the maze was only a collection of walls. ",
"To create the base of the maze, a separate circle was added and sized to fit the outer wall of the maze. ",
"The object to path option was selected for the base only to make it a solid object. ",
"The walls and the base were overlaid so that all edges were overlapping. ",
"Following this, both the base and the walls of the maze were selected (ctrl-A), then under the extensions toolbar, the \"generate from path\" option was selected and the paths to openSCAD option was chosen to convert to the two objects into an openSCAD file (.scad).",
"\n\nThe openSCAD file was opened in OpenSCAD v.2015.03 (free software released under the general public license version 2). ",
"OpenSCAD is able to render the .scad file into a 3D image. ",
"In this case, the file still had the two parts overlaid in space, the \"poly_path,\" the base of the object and the \"poly_line,\" the maze wall portion of the object. ",
"The file was first previewed to confirm that the two parts of the image overlapped. ",
"If not, the file was created again in Inkscape to ensure that the edges aligned. ",
"Initially, the image appeared as a solid cylinder as both the base and the walls of the image were fully extruded to the same height by default. ",
"The extrusion height of both objects was modified in the left hand panel by finding the section of code related to the height of each object (end of the code) and revising the poly_path (base) height to 1 mm from the bottom of the image, and poly_line (wall) 6 mm from the bottom, or 5 mm above the base (6 mm--1 mm). ",
"The refreshed image showed a 1 mm base under a 5 mm wall. ",
"When the maze was ready, the image was rendered and saved as an .stl file for printing. ",
"We used UP! ",
"version 2.18 (Beijing TierTime Technology Co. Ltd.) to print the maze files using an UP mini 3D printer with white ABS filament (3D printing systems Pakuranga, Auckland, New Zealand). ",
"A piece of clear 2 mm thick Perspex was attached to the top of the maze with masking tape so that fly behaviour could be observed.",
"\n\n{#insects-11-00256-f001}\n\n{#insects-11-00256-f002}\n\n{#insects-11-00256-f003}\n\n{#insects-11-00256-f004}\n\ninsects-11-00256-t001_Table 1\n\n###### \n\nTime for the first female and male *D. simulans* to find each other in a maze with or without a pseudoscorpion predator (*C. cancroides*) present and the percentage of flies finding each other within a treatment in trials with varying fly density.",
"\n\n No. ",
"of Pairs Predator Present Mean Initial Discovery Time \\% of Trials with Successful Mate Finding \\% of Trials with Mating Occurring Percentage of Flies Mating Number of Flies Killed (In N Replicates)\n -------------- ------------------ ----------------------------- ------------------------------------------- ------------------------------------ ---------------------------- ------------------------------------------\n 1 Yes 25m15s 53% 3% 3% 8 (8)\n 1 No 17m10s 76% 10% 10% \\-\n 3 Yes 16m35s 96% 43% 15% 12 (11)\n 3 No 16m31s 96% 40% 20% \\-\n 5 Yes 12m40s 100% 46% 13% 13 (10)\n 5 No 7m28s 100% 60% 25% \\-\n\ninsects-11-00256-t002_Table 2\n\n###### \n\nGLM Poisson log-link regression of time taken for a pair of *D. simulans* to find each other (first pair only), with or without a *C. cancroides* (predator) present. ",
"The reference parameters are one pair of flies and no predator present. ",
"The antilog can be considered as the multiple by which the change has occurred compared to the expected result for the reference parameters (1 pair and no predator).",
"\n\n Parameter Estimate (s.e.) ",
" t *p* Antilog of Estimate\n ------------------ ------------------ -------- --------- ---------------------\n 3 pairs −0.2372 (0.0480) −4.95 \\<0.001 0.7888\n 5 pairs −0.7345 (0.0540) −13.61 \\<0.001 0.4797\n Predator present 0.2651 (0.0413) 6.41 \\<0.001 1.303\n\ninsects-11-00256-t003_Table 3\n\n###### \n\nGLM binomial logit-link regression of the probability that mating would occur at all in trials where different numbers of pairs of *D. simulans* were in the presence of a predator (*C. cancroides*) or not. ",
"The reference parameters were one pair of flies and no predator present. ",
"The antilog can be considered as the multiple by which the change has occurred compared to the expected result for the reference parameters (one pair and no predator).",
"\n\n Parameter Estimate (s.e.) ",
" t *p* Antilog of Estimate\n ------------------ ----------------- ------- --------- ---------------------\n 3 pairs 1.844 (0.538) 3.43 \\<0.001 6.324\n 5 pairs 2.095 (0.530) 3.96 \\<0.001 8.127\n Predator present −0.167 (0.259) −0.64 0.519 0.8463\n"
] | {
"pile_set_name": "PubMed Central"
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[
"// Copyright (c) Lawrence Livermore National Security, LLC and other VisIt\n// Project developers. ",
" See the top-level LICENSE file for dates and other\n// details. ",
" No copyright assignment is required to contribute to VisIt.",
"\n\n// ************************************************************************* //\n// File: avtTransformFilter.h\n// ************************************************************************* //\n\n#ifndef AVT_Transform_FILTER_H\n#define AVT_Transform_FILTER_H\n\n\n#include <avtPluginFilter.h>\n#include <avtSingleFilterFacade.h>\n#include <TransformAttributes.h>\n\nclass avtLinearTransformFilter;\nclass avtSimilarityTransformFilter;\nclass avtCoordSystemConvert;\n\n\n// ****************************************************************************\n// Class: avtTransformFilter\n//\n// Purpose:\n// A plugin operator for Transform.",
"\n//\n// Programmer: Jeremy Meredith\n// Creation: September 24, 2001\n//\n// Modifications:\n//\n// Kathleen Bonnell, Wed Nov 28 16:59:53 PST 2001\n// Added UpdateDataObjectInfo, ModifyContract.",
"\n//\n// Hank Childs, Tue Mar 5 16:05:25 PST 2002\n// Removed UpdateDataObjectInfo, since what is was doing is also\n// applicable to the base class, so I pushed the logic into it.",
"\n//\n// Kathleen Bonnell, Thu Apr 10 11:07:48 PDT 2003 \n// Store inverse matrix for possible use later in the pipeline. ",
"\n// Added PostExecute method.",
"\n// \n// Hank Childs, Tue Jul 1 08:59:08 PDT 2003\n// Moved original avtTransformFilter to libpipeline with the name\n// avtSimilarityTransformFilter. ",
" Re-worked this filter to be a single\n// filter facade.",
"\n//\n// Hank Childs, Tue Feb 1 16:37:56 PST 2005\n// Allow for coordinate system transformations as well.",
"\n//\n// Jeremy Meredith, Tue Apr 15 13:17:24 EDT 2008\n// Added linear transform.",
"\n//\n// Tom Fogal, Tue Jun 23 20:53:28 MDT 2009\n// Added const version of GetFacadedFilter.",
"\n//\n// ****************************************************************************\n\nclass avtTransformFilter : public virtual avtPluginFilter,\n public virtual avtSingleFilterFacade\n{\n public:\n avtTransformFilter();\n virtual ~avtTransformFilter();\n\n static avtFilter *Create();\n\n virtual const char *GetType(void) { return \"avtTransformFilter\"; };\n virtual const char *GetDescription(void)\n { return \"Transforming\"; };\n\n virtual void SetAtts(const AttributeGroup*);\n virtual bool Equivalent(const AttributeGroup*);\n\n protected:\n TransformAttributes atts;\n avtLinearTransformFilter *ltf;\n avtSimilarityTransformFilter *stf;\n avtCoordSystemConvert *csc;\n\n virtual avtFilter *GetFacadedFilter(void);\n virtual const avtFilter *GetFacadedFilter(void) const;\n\n static void UpdateDataObjectInfoCB(avtDataObject_p &input,\n avtDataObject_p &output, void *);\n};\n\n#endif\n"
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[
"Pojat ovat poikia, kuuluu ikivanha sanonta.",
"\n\nPoikuuteen ja miehisyyteen katsotaan yleisesti kuuluvan tietynlaiset asiat. ",
"Poikien odotetaan olevan maskuliinisia, rationaalisia, urheilullisia, kovia ja uhkarohkeitakin. ",
"Väkivaltakin on poikien juttu.",
"\n\nKaikki ominaisuudet asteikon toisessa päässä ovat tyttöjen tai \"homojen hommaa\".",
"\n\nMutta kaikki pojat eivät mahdu yhteiskunnan heille muovaamaan ahtaaseen muottiin.",
"\n\nOn arvokkaampaa hoivata konetta kuin elävää ihmistä\n\n\"Sukupuoli \"lukitaan\" viimeistään synnytyssairaalassa värikoodattujen tunnisteiden ja sosiaaliturvatunnuksen myötä. ",
"Elämänkulun myöhemmissä vaiheissa biologisiksi pojiksi tunnistettuja lapsia ja nuoria ympäröi maailma, jossa heiltä odotetaan poikamaiseksi katsottua käytöstä.\"",
"\n\nLaura Koivunen / Yle Uutisgrafiikka\n\nNäin todetaan juuri julkaistun Poikatutkimus-kirjan johdannossa. ",
"Kirja on ensimmäinen suomalaista poikatutkimusta esittelevä tieteellinen kokoomateos.",
"\n\nYksi kirjan tavoite on tarkastella kriittisesti yhteiskunnan sukupuolistuneita rakenteita, kertoo yksi kirjan toimittajista, Oulun yliopiston akatemiatutkija Tuija Huuki.",
"\n\nHuukin mukaan maskuliinisuutta ja sitä, mitä pidetään miehisenä, on arvostettu aina korkeammalle. ",
"Rationaalisuus on arvokkaampaa kuin tunnelähtöisyys.",
"\n\n– Saat isompaa palkkaa siitä, kun painat paperitehtaalla nappia ja huolehdit koneesta, kuin siitä, että olet päiväkodissa ja huolehdit elävästä ihmisestä. ",
"Kyse lienee paitsi logiikasta, jossa tuottavuus määrittää työntekijöiden palkkaa melko paljon, myös siitä, että miehisyyttä ja miehiseksi katsottuja asioita on arvostettu ja arvostetaan enemmän.",
"\n\nSiksi miehiseksi katsotuista asioista poikkeaminen on pojalle usein vaikeaa. ",
"Tyttöjen poikamaista toimintaa ymmärretään paremmin, mutta poikien tyttömäistä toimintaa ei välttämättä katsota kovin hyvällä.",
"\n\nTyöelämässä ja maailmassa tarvitaan sekoittumista\n\n\"Kaksinapaisen tyttö-poika -jaottelun käyttäminen kaiken selittävänä \"supermuuttujana\" on syytä kyseenalaistaa.\" (",
"ote Poikatutkimus-kirjasta)\n\nMiksi?",
"\n\nSiksi, että se kaventaa ihmisessä olevia moninaisia kykypotentiaaleja, perustelee Tuija Huuki. ",
"Ihmisen pitäisi pystyä hallitsemaan kaikenlaisia taitoja hiljentymisestä ja itkemisestä voimalliseen itseilmaisuun. ",
"Kaikille taidoille on paikkansa maailmassa ja elämässä jossain kohtaa.",
"\n\nJos poikia kielletään ottamasta haltuun intiimi herkkyys ja tunteet ja tytöiltä evätään rationaalisuus ja johtajuus, jää puolet ihmisessä olevista kykypotentiaaleista käyttämättä.",
"\n\n– Poikien pitäisi pystyä ottamaan haltuun niitä ominaisuuksia, mitä pidetään tyttömäisinä, koska niitä tarvitaan elämässä, Huuki sanoo.",
"\n\nHuukin mielestä on vahingollista, jos poika ei opi suremaan ja kohtaamaan toisia tunnetasolla. ",
"Tunteiden patoaminen voi johtaa siihen, että ennemmin tai myöhemmin paineet kasvavat liian suuriksi ja tunteet purkautuvat ulos hallitsemattomasti, esimerkiksi päihteiden käytöllä tai väkivallalla.",
"\n\n– Elämä on aika kuluttavaa, jos pitää padota tunteet, tehdä riskialttiit temput, ryypätä ja rellestää osoittaakseen olevansa mies.",
"\n\nOulun yliopiston akatemiatutkija Tuija Huuki. ",
"Mikko Törmänen\n\nHuukin mukaan työelämässä ja maailmassa tarvitaan sekoittumista. ",
"Hoiva-aloille tarvitaan miehiä ja miesenergiaa ja johtajaportaaseen naisia ja naisenergiaa.",
"\n\n– Tiedämme mitä ylikorostunut teknologiakeskeisyys ja rationaalisuus ovat tuottaneet planeetallemme. ",
"Sieltä ovat puuttuneet herkkyys ja hoiva sekä vastuuajattelu, jonka pitäisi kohdistua myös ympäristöön. ",
"Ja tässä on tulos.",
"\n\nMieheys on jatkuvassa muutoksessa\n\nMiehenä olemista ja miehen mallia perustellaan usein hormoneilla tai evoluutiolla. ",
"Huukin mukaan tutkimuksen valossa väite on liian yksioikoinen.",
"\n\n– Evoluutio ei yksin selitä miehenä toimimista ja mieheksi kasvamista. ",
"Tutkimus ei ole pystynyt osoittamaan selkeää syy-seuraus -suhdetta luolamiesajoilta. ",
"Se on spekulatiivista.",
"\n\n– Kirjan sanoma on se, että mieheys rakentuu lukemattomien eri tekijöiden vaikutuksesta. ",
"Siinä on niin paljon kulttuurisesti ja sosiaalisesti rakentunutta, että niitä asioita voidaan muuttaa. ",
"Ja mieheys muuttuukin, koko ajan.",
"\n\nHuuki antaa esimerkkejä historiasta. ",
"Antiikin Kreikassa homoseksuaalisuus oli normaalia. ",
"Barokin puuteroidut miehet olivat todella laittautuneita ja pröystäileviä sukkahousuissaan ja peruukeissaan.",
"\n\nMuutoksen mahdollisuus on Huukin mukaan koko ajan olemassa. ",
"Pojat ovat kyvykkäitä vaikuttamaan omiin ympäröiviin olosuhteisiinsa.",
"\n\nMonipuolisempia uravalintoja ja lisää lukuintoa\n\n\"Kun osoitetaan, että poikia on monenlaisia ja poikuus on monenlaista - ja kun samalla puretaan sukupuolistuneita käytäntöjä - avautuu merkittäviä mahdollisuuksia myönteisiin yhteiskunnallisiin seurauksiin.\"",
"\n\nPoikatutkimus-kirjan viesti on, että ahtaiden sukupuolinormien ylittäminen on mahdollista ja se kannattaa.",
"\n\nEnsinnäkin se voisi toimia ratkaisuna oppimisen ongelmiin, kasvattaa koulumyönteisyyttä ja lisätä poikien lukuintoa.",
"\n\nHuukin mukaan lukutoukkapoika nähdään usein ei-poikamaisena. ",
"Eräässä tutkimuksessakin poika kertoi, että yleensä vain homot pitävät lukemisesta. ",
"Lukeminen voi siis joissain poikayhteisöissä olla huono juttu ja heikentää pojan asemaa.",
"\n\nHuukin mukaan ilmiötä ei aina tunnisteta.",
"\n\n– Vaikka me kuinka kehittelemme lukumyönteistä pedagogiikkaa ja kannustamme lukemaan, se ei välttämättä kohtaa poikien omia alakulttuurisia arvoja. ",
"Meidän pitäisi pureutua niihin.",
"\n\nLaveampi mieskuva toisi mukanaan myös monipuolisempia uravalintoja ja vähemmän sukupuolistuneet työmarkkinat.",
"\n\nJos pojat pienestä pitäen kasvatetaan ajattelemaan, että heitä varten ovat vain matematiikka ja tekniikka, silloin he myös hakeutuvat sellaisille aloille, Huuki perustelee.",
"\n\nSama koskee myös tyttöjä, mutta ei yhtä paljon.",
"\n\n– Siihen on satsattu, että tytöt hakeutuisivat teknisille aloille. ",
"Mutta siihen ei ole vielä juurikaan satsattu, että pojat hakeutuisivat esimerkiksi hoiva-aloille, koska miehen feminiinisyys on tiedostamattomalla tasolla epäarvo.",
"\n\nLaura Koivunen / Yle Uutisgrafiikka\n\nVähemmän naisiin kohdistuvaa väkivaltaa\n\nMitä väkivaltaisempi mieskuva on, sitä enemmän on myös naisiin kohdistuvaa väkivaltaa.",
"\n\nHuuki kertoo, että kulttuuriantropologiassa on dokumentoitu eri aikoina maailman eri kolkissa kulttuureja, joissa esiintyy hyvin vähän väkivaltaa.",
"\n\nJa toisaalta on maita, joissa sitä on paljon, kuten Suomessa.",
"\n\n– Meillähän on kovin karski mieskuva, jos vertaa vaikka Ruotsiin. ",
"Ruotsissa maskuliinisuuskuva on pehmeämpi kuin meillä ja tämä voi olla yksi syy siihen, miksi meillä monesti kuulee puhuttavan siitä, miten ruotsalaismiehet ovat homoja. ",
"Vaikka otaksuttavasti ne homoseksuaalisuuden luvut eivät ole siellä yhtään sen kummempia kuin täälläkään.",
"\n\n– Jos meidän mieskuva muuttuu pehmeämpään suuntaan, sen seurauksena naisiin kohdistuva väkivalta todennäköisesti vähenee. ",
"Miehet pystyvät käsittelemään tunteitaan muutenkin kuin lyömällä vaimoa, Huuki sanoo.",
"\n\nPojat ovat muutakin kuin poikia\n\nHuukin mukaan tähänastinen keskustelu pojista on ollut liian yksisilmäistä: pojat on niputettu yhdeksi ryhmäksi.",
"\n\nKeskustelussa ei ole Huukin mielestä päästy riittävän yksityiskohtaiseen analyysiin siitä, keitä poikia tarkemmin ottaen koulutuksen, lukemisen, väkivallan ja syrjäytymisen haasteet oikein koskevat. ",
"Keskustelu on perustunut olettamukselle, että poikia kohtaavat ongelmat koskevat kaikkia poikia samalla tavalla.",
"\n\n– Meillähän on ihan valtavasti hyvin pärjääviä poikia. ",
"Jos katsomme pitkittäisesti, mitä tapahtuu, kun pojat tulevat nuoreen aikuisuuteen, niin nimenomaan nuoret miehethän nappaavat johtavat paikat työelämässä esimerkiksi. ",
"Se, jos mikä, on pärjäämisen merkki.",
"\n\nSukupuolentutkimuksessa ja Poikatutkimus-kirjassa ei pyritä yleistämään asioita vaan katsotaan, mitä kaikkea poikuus on ja miten moninaisessa maailmassa pojat elävät.",
"\n\nTyttötutkimusteoksia on julkaistu Suomessa jo kuusi. ",
"Poikatutkimus-kirjalle on siis tilausta paitsi yhteiskunnallisesti myös suomalaisen tieteen kentällä. ",
"Kirjassa on useita tieteellisiä artikkeleita, jotka käsittelevät muun muassa miehen mallin käsitettä historian valossa, poikien koulumaailmaa ja lukemista, poikien haavoittuvuutta, väkivaltaista maskuliinisuutta sekä poikien erilaisia yhteisöjä.",
"\n\nKirja kokoaa samojen kansien väliin jatkuvasti kehittyvää sukupuolivastuullista poikatutkimusta. ",
"Sukupuolivastuullisuus tarkoittaa muun muassa sitä, että tutkijat ymmärtävät, että on monenlaisia poikia ja myös sen, millaisia sukupuolistuneita odotuksia poikiin kohdistuu.",
"\n\nEsimerkiksi työväenluokkaisista taustoista tulevat pojat kasvavat merkittävästi erilaisessa kokemusmaailmassa kuin keskiluokkaisista taustoista tulevat pojat.",
"\n\n– Tämän päivän poikatutkimus analysoi sukupuolta ja sukupuolen sisäisiä eroja. ",
"Ei tuijoteta vain sukupuolten välisiä eroja, vaan ymmärretään, että tytöissä ja pojissa on paljon samanlaisuuksia. ",
"Ja sukupuolen sisällä, monilla elämänalueilla, on niin paljon eroja, että ne usein ylittävät sukupuolten väliset erot.",
"\n\nLue myös:\n\n\"Poikien nokittelukulttuurissa on vaikea olla oma itsensä\" – Käytännön vinkkejä kasvattajille siitä, miten \"poikakoodi\" puretaan\n\nRiku Rantala: Miehuus nousee pelosta, ja tätä naiset eivät ymmärrä\n\n“Olenko minä ok, riitänkö minä ja onko maailma tällainen?” – ",
"Liian monella pojalla ei edelleenkään ole tilaa puhua tärkeistä asioista"
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"Plasma antioxidants: evidence for a protective role against reactive oxygen species following cardiac surgery.",
"\nTotal plasma antioxidant status (TPAS), lipid peroxide concentration (LPX) and cardiac troponin T (cTnT) were measured in 24 patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB). ",
"Samples were obtained preoperatively and at 1.5 h, 6 h, 24 h and 72 h after CPB. ",
"The absolute TPAS values were significantly lower at 1.5 h, 6 h, 24 h and 72 h after CPB than were preoperative values (P < 0.05). ",
"The LPX concentration was significantly elevated at 1.5 h after CPB (P < 0.05). ",
"Cardiac troponin T concentrations were significantly elevated at all time points postoperatively (P < 0.05). ",
"Preoperative TPAS values were significantly correlated with the magnitude of fall in TPAS at 1.5 h (P < 0.05). ",
"The greater the fall in TPAS between 0 and 1.5 h, the less LPX was formed between 0 and 1.5 h. The LPX at 1.5 h displayed a significant correlation with cTnT release from myocardial myocytes (P < 0.05). ",
"These data provide evidence for the first time that the consumption of antioxidants during CABG surgery with CPB protects against the production of reactive oxygen species and subsequent myocyte necrosis. ",
"Furthermore, the availability of protective antioxidants is dependent upon preoperative TPAS."
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"News\n\nMetro passengers have voted in their thousands and have chosen a vibrant light blue colour scheme for the new-look Central Station in Newcastle.",
"\nMore than 5,000 people voted after Nexus gave passengers a chance to have their say on the new colour scheme.",
"\nNexus, which owns, manages and is...\n\nNexus, which owns, manages and is modernising the Tyne and Wear Metro, has installed automatic ticket gates at Chichester Metro station in South Tyneside.",
"\nThe new ticket gates have been installed over the last two weeks and will be switched on in February.",
"\nIt will mean that the station will be...\n\nNexus, which owns and manages the Tyne and Wear Metro, has given its new apprentices the chance to get first-hand experience of operating multi-million pound railway maintenance equipment.",
"\nTen first year apprentices, who are part of the three-year Nexus Apprenticeship Scheme, got the chance to...\n\nNexus, the public body which owns and manages the Tyne and Wear Metro, has donated a set of level crossing stop lights to one of the North’s most well-known heritage railways.",
"\nThe Wensleydale Railway in North Yorkshire, which is run by volunteers, has been given a used set of red stop lights from...\n\nA Metro driver who was involved in a fatal collision while in the cab of his train has trained as a counsellor to help colleagues who find themselves in the same position.",
"\nAlan Hudson, a Metro driver for 14 years, decided to become a counsellor as a way of coping with the trauma of an incident five...\n\nMetro passengers who buy annual MetroSavers can now buy tickets priced zone-by-zone for annual season tickets, offering cheaper travel for thousands of commuters.",
"\nThe zonal MetroSavers are now on sale exclusively from all Nexus Travelshops. ",
"They will be available from all Metro station ticket...\n\nNexus, which owns and manages the Tyne and Wear Metro, has embarked on a third consecutive year of apprentice recruitment for its rail engineering section.",
"\nA total of ten apprentices are being sought to join the Nexus Apprenticeship Scheme in the autumn of 2015. ",
"Nexus expanded the scheme in 2013...\n\nCustomer service workers at Nexus are playing a key role in keeping public transport hubs safe from sex offenders.",
"\nThe Nexus staff have appeared in a new national training video, commissioned by the independent charity Leisurewatch, on how to spot the characteristics and actions of potential...\n\nNexus, which owns and manages the Tyne and Wear Metro, has successfully completed an £800,000 project to improve safety at Metro level crossings – including the introduction of warning lights for pedestrians.",
"\nFive level crossings, which are maintained by Nexus, have received a series of safety...\n\nEmployers are signing up for a staff travel scheme on Tyne and Wear Metro in record numbers after the Corporate MetroSaver ticket scheme was extended earlier this year, allowing smaller companies with fewer staff to sign up.",
"\nAs a result, Nexus has seen a rise in the number of enquiries and..."
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"The investigators will test the hypothesis that pulsatile testosterone delivery which emulates physiologically episodic secretion of androgen will inhibit LH and FSH release to a greater extent than continuous infusion of the same mass of testosterone over 24 hours in men with blocked Leydig-cell androgen biosynthesis. ",
"In young men, continuous testosterone suppresses LH more than intermittent. ",
"The study is being extended to older men."
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"Finally saw Skyfall\nIf we take bond's and Javier Bardem's performance and the pretty bond girl out of the movie, we are left with a steamy pile of sh*t.",
"\nThere are more holes in the plot than a pasta strainer"
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"All relevant data are within the paper and its Supporting Information files.",
"\n\nIntroduction {#sec001}\n============\n\nOligodendrocytes possess a pivotal role in ensheathing axons by myelin. ",
"Their pathology contributes to various disorders including multiple sclerosis (MS), white matter stroke, traumatic CNS injury, and age-related myelin loss. ",
"Replacing the oligodendrocytes is a promising approach for therapeutic applications, in particular the generation of autologous sources of oligodendrocyte progenitor cells (OPCs) is of great interest.",
"\n\n*In vivo* conversion of somatic cells into desired cell types is considered a proper approach to generate progenitor repair cells for therapeutic purposes with emphasis on organs that have a low capacity to regenerate. *",
"In vivo* lineage reprogramming approaches have been demonstrated in the brain\\[[@pone.0203785.ref001], [@pone.0203785.ref002]\\], spinal cord \\[[@pone.0203785.ref003]\\], heart \\[[@pone.0203785.ref004]\\], and pancreas \\[[@pone.0203785.ref005]\\].",
"\n\nAstrocytes are the most plentiful cellular components of glial scars which develop following neural cell loss from degenerative diseases and traumatic injuries by reactivation of astrocytes and their secretions \\[[@pone.0203785.ref006]\\]. ",
"Previous studies demonstrated that astrocytes could be directly converted into neurons or stem-like cells by the forced expression of transcription factors in vitro \\[[@pone.0203785.ref007]--[@pone.0203785.ref011]\\], which highlights the capability of fate change of these somatic glial cells. ",
"Recently, several attempts have been made to convert astrocytes within the brain parenchyma to neurons by Sox2 \\[[@pone.0203785.ref003], [@pone.0203785.ref012], [@pone.0203785.ref013]\\], NeuroD1 \\[[@pone.0203785.ref001]\\], Ascl1 \\[[@pone.0203785.ref014]\\], and MicroRNA 302/367 \\[[@pone.0203785.ref015]\\]. ",
"In this strategy, instead of surgical removal of the glial scar, reactive astrocytes are converted into progenitor cells that contribute to tissue repair. ",
"Developmentally, astrocytes and oligodendrocytes are produced from glial progenitors and may be considered as differentiated cells with similar epigenetic states \\[[@pone.0203785.ref016]\\]. ",
"In previous reports conversion of astrocytes to myelinating cells was done using MicroRNA 302/367 and transcription factor Oct4 that were not specific to oligodendrocytes \\[[@pone.0203785.ref017], [@pone.0203785.ref018]\\]. ",
"The same MicroRNA also produced neuroblasts which had the capability to differentiate into mature neuron-like cells in normal brains \\[[@pone.0203785.ref015]\\] and animal models of neurodegeneration and Alzheimer's disease \\[[@pone.0203785.ref019], [@pone.0203785.ref020]\\]. ",
"Therefore, in the current study, we attempted to locate a single transcription factor specific to oligodendrocyte lineage cells that had the capability for in vivo conversion of astrocytes into OPCs.",
"\n\nSox10 is a transcription factor related to the sex determining region Y (SRY)-boxes gene family expressed constantly throughout OPC development into mature oligodendrocytes\\[[@pone.0203785.ref021]\\]. ",
"Numerous evidences suggest that Sox10 is as a master regulator in the developmental process of oligodendrocytes and activation of myelination genes \\[[@pone.0203785.ref022]--[@pone.0203785.ref024]\\]. ",
"This transcription factor in combination with NKX6.2 and Olig2 \\[[@pone.0203785.ref025]\\]or Olig2 and ZFP536 \\[[@pone.0203785.ref026]\\] has the capability to reprogram rodent fibroblasts into induced OPCs (iOPCs). ",
"Continuous expression of Sox10 in satellite cells of dorsal root ganglion via a transgene approach has led to ectopic development of oligodendrocytes \\[[@pone.0203785.ref027]\\]. ",
"While NFIA is known as a key astrocyte fate determinant factor, it has been reported that Sox10 suppresses astrocytic differentiation by an antagonistic relationship with NFIA and regulating the oligodendrocyte and astrocyte fate during development \\[[@pone.0203785.ref028]\\] In fact, Sox10 positive astrocytes have been hardly detected in corpus callosum and brain cortices \\[[@pone.0203785.ref029]\\].This reports also provide another explanation for using Sox10 to convert astrocytes to OPCs.",
"\n\nTo the best of our knowledge, this is the first report of using forced expression of a myelination specific transcription factor, Sox10 that has induced astrocytes to reprogram into oligodendrocyte lineage cells.",
"\n\nMaterials and methods {#sec002}\n=====================\n\nViral particle preparation {#sec003}\n--------------------------\n\nThe desired construct containing a CDS for the Sox10 gene was generated by ligation between a sox10 CDs (extracted from Tet-O-FUW-Sox10) and a pSSFV-IRES-GFP plasmid, to acquire the pSSFV-Sox10-IRES-GFP. ",
"Tet-O-FUW-Sox10 was a gift from Marius Wernig (Addgene plasmid \\# 45843). ",
"The schematic diagram for vectors are presented in Panel A in [S1 Fig](#pone.0203785.s001){ref-type=\"supplementary-material\"}.",
"\n\nConfluent human embryonic kidney cells (HEK 293T) were transfected with pSSFV-*Sox10*-IRES-*GFP* or pSSFV-IRES-*GFP* constructs and packaging plasmids (pCMV-vsvg and pCMV-gp) using Lipofectamine 3000 (Life Technologies). ",
"At 48 and 72 h post-transfection the cell culture media was collected, filtered, and centrifuged to concentrate the viral particles. ",
"The collected particles were re-suspended in 100 μL PBS for subsequent infections.",
"\n\nInduction of demyelination in animals {#sec004}\n-------------------------------------\n\nMale C57BL/6 mice, 7--8 weeks old, (Royan Institute) were housed in a climate-controlled room with access to food and water ad libitum. ",
"All experiments were performed in accordance with the international guide for the care and use of laboratory animals. ",
"The experimental procedures wereevaluated and approved by the Committee for Ethics in Animal Research at Royan Institute. ",
"Attempts were made to minimize the number of animals used and their suffering. ",
"For demyelination induction, mice were fed with 0.2% (w/w) oxalic acid bis(cyclohexylidenehydrazide) (cuprizone; Sigma-Aldrich, C9012) mixed into their normal chow for 12 weeks. ",
"We conducted the Y-maze test and histological studies to confirm the presence of behavioral impairment due to demyelination and extensive demyelination in their brains.",
"\n\nSpontaneous alternations within the Y-maze was tested prior to induction of demyelination and after the 12-week period of cuprizone administration in their chow. ",
"The Y-maze arms were marked A, B, and C. The number of overlapping entrance sequences (e.g., ABC, BCA) was defined as the number of spontaneous alternations. ",
"We calculated the percent of alternations as follows \\[[@pone.0203785.ref030]\\].",
"\n\nPercent of alternations = (number of alternations)/ (total number of arm entries-2) ×100.",
"\n\nThe number of total arm entries was calculated as the index of ambulatory activity in the Y-maze. ",
"We excluded any animals with scores of less than six entries.",
"\n\nAnimal interventions {#sec005}\n--------------------\n\nWe evaluated the possible conversion of transduced cells into myelinating cells. ",
"The schematic diagram for animal interventions are presented in Panel B in [S1 Fig](#pone.0203785.s001){ref-type=\"supplementary-material\"}. ",
"Mice with cuprizone induced demyelination were anesthetized with ketamine (100 mg/kg, i.p., ",
"GE Healthcare, Germany) and xylazine (5 mg/kg, i.p., ",
"Bayer, Germany), then placed in a stereotaxic frame in a flat skull position. ",
"Lentiviral particles (2 μl of concentrated solution) were injected into each corpus callosum at the following coordinates from the atlas of Paxinos and Watson: anteroposterior (AP) = -1.06, mediolateral (ML) = 1 from the bregma, and dorsoventral (DV) = 1.5 mm from the skull. ",
"Animals received a normal diet for 3 or 8 weeks after the injection.",
"\n\nWe assessed the conversion of astrocytes to oligodendrocyte lineage cells by transducing purified astrocytes (see below) with pSSFV-Sox10-IRES-GFP or pSSFV-IRES-GFP viruses. ",
"After 48 h, we transplanted 1×10^5^ cells into the corpora callosa of the mice with demyelinated brains. ",
"The animals received immunosuppression by daily injections of cyclosporine A (20 mg/kg; Novartis Pharmaceuticals, East Hanover, NJ, USA) from 2 days before surgery until 3 weeks after transplantation.",
"\n\nTissue sampling and immunohistofluorescence (IHF) studies {#sec006}\n---------------------------------------------------------\n\nAnimals were deeply anesthetized and perfused transcardially with saline, followed by 4% paraformaldehyde (Sigma-Aldrich, P6148) prepared in PBS. ",
"Then, the brains were removed, post-fixed overnight in 4% PFA, and cryoprotected for 48 h in 30% sucrose. ",
"Samples were embedded in optimal cutting temperature (OCT) compound and serial 10 μm-thick coronal sections were cut using a cryostat (Leica Microsystems, CM 1850). ",
"The cut sections were stored at -20°C and then processed for immunohistofluorescence (IHF) analysis.",
"\n\nSections were washed three times in PBS, then incubated in blocking solution that consisted of 0.1% Triton X-100 (Sigma-Aldrich, T8532), 1% bovine serum albumin, and 5% normal serum in PBS for 1 h at room temperature, followed by an overnight incubation with primary antibodies against NG2, Olig2, glial fibrillary acidic protein (Gfap), PdgfRα, Plp, Mbp or NeuN at 4°C. ",
"Sections were washed three times with PBS and incubated at room temperature for 2 h with the secondary antibody. ",
"List of antibodies is provided in [S1 Table](#pone.0203785.s005){ref-type=\"supplementary-material\"}. ",
"After treatment with 4′, 6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich; D-8417), the sections were again washed three times with PBS. ",
"For myelin staining with FluoroMyelin, sections were incubated with FluoroMyelin Red (Invitrogen F34652, 1/300) solution dissolved in PBS for 20 minutes at room temperature. ",
"Sections were coverslipped and then Images were obtained on an Olympus IX51 fluorescence microscope with a DP72 digital camera and analySIS LS Starter software version 3.2.",
"\n\nAstrocyte isolation and culture {#sec007}\n-------------------------------\n\nAstrocytes were isolated based on the procedure by Schildge et al. ",
"\\[[@pone.0203785.ref031]\\] with some modifications. ",
"After removal of the meninges, grey matter tissue from the cerebral cortexes of P3-P5 C57BL/6J mice was dissected and mechanically dissociated. ",
"The suspension was centrifuged for 5 min at 1000 rpm, re-suspended, and plated in a medium that contained DMEM/F12 (Invitrogen), 3.5 mM glucose (Sigma), 10% fetal bovine serum (Gibco), and 1% penicillin/streptomycin (Gibco). ",
"After 24 h, the contaminating oligodendrocyte precursor cells were removed by vigorous manual agitation. ",
"One week later, cells were passaged using and plated in fresh medium. ",
"The CD44+ cell population was purified from a primary culture of astrocytes by incubation with anti-CD44 antibody (BD550538) followed by MACS selection (Miltenyi Biotech, Germany). ",
"We confirmed the fate of extracted cells and the purity of the sorted astrocytes by immunocytofluorescence ([S2](#pone.0203785.s002){ref-type=\"supplementary-material\"} and [S3](#pone.0203785.s003){ref-type=\"supplementary-material\"} Figs).",
"\n\nTransduction of astrocytes in culture {#sec008}\n-------------------------------------\n\nPurified astrocytes were transduced with prepared viral particles and kept in astrocyte media for an additional 48 h. Afterwards, transduced cells were seeded on 0.1 mg/ml poly-l-ornithine (Sigma-Aldrich; P4707) and 10 μg/ml laminin (Sigma-Aldrich; L2020) for in vitro induction or transplanted into animal brains for in vivo transdifferentiation. ",
"In culture, the induction medium was permissive to oligodendrocyte fate acquisition and consisted of DMEM/F12 (Invitrogen) supplemented with N2 (Invitrogen), B27 without vitamin A (Invitrogen), 2 mM Glutamax (Invitrogen), 200 ng/ml SHH (Royan Biotech), 20 ng/ml bFGF (Royan Biotech), and 20 ng/ml PDGF-AA (Royan Biotech). ",
"Cultures were fixed after 21 days and stained against the OPC markers, Olig2 and NG2, and neuronal marker Tuj1; after which we determined the percent of transduced (GFP+) cells positive for NG2, Olig2 and Tuj1.",
"\n\nData analysis {#sec009}\n-------------\n\nWhen required, we performed cell counts in several randomly selected views of all samples from each brain section. ",
"At least 8 sections were counted per animal and each animal group included 3 mice. ",
"For statistical comparison of the means of different groups, one-way ANOVA followed by the Tukey post-test was used except for the Y-maze data which we compared with the Wilcoxon signed-rank test for matched pairs. ",
"The results were expressed as mean±SEM. ",
"A probability of p\\<0.05 was considered statistically significant.",
"\n\nResults {#sec010}\n=======\n\nPrevious studies reported that a 12-week cuprizone diet led to extensive demyelination in the mice brains. ",
"This finding was particularly noted in the corpus callosum of each brain. ",
"Researchers have reported that a 12-week cuprizone diet decreased endogenous remyelination \\[[@pone.0203785.ref032], [@pone.0203785.ref033]\\] and therefore provides an adequate model to study the capability of *in situ* induced OPCs or transplanted cells for myelination. ",
"Astrogliosis, defined as astrocyte hypertrophy and reactivity, is one of the most important features of this demyelination model \\[[@pone.0203785.ref034]\\]. ",
"Gfap staining showed extensive astrocyte reactivation within the brains ([Fig 1A](#pone.0203785.g001){ref-type=\"fig\"}). ",
"FluoroMyelin staining showed extensive corpus callosum demyelination in cuprizone-treated animals compared to control mice of the same age (20 weeks; [Fig 1B](#pone.0203785.g001){ref-type=\"fig\"}). ",
"Demyelination was also confirmed by luxol fast blue staining ([Fig 1C](#pone.0203785.g001){ref-type=\"fig\"}). ",
"Spatial working memory is affected following cuprizone induced demyelination in mice \\[[@pone.0203785.ref035]\\]. ",
"Measurements of spontaneous alternations within the symmetric Y maze confirmed the presence of myelination decline in the study mice. ",
"The treated mice showed significant impairment in terms of total arm entries and spontaneous alternations compared to baseline ([Fig 1D and 1E](#pone.0203785.g001){ref-type=\"fig\"}).",
"\n\n{#pone.0203785.g001}\n\nWe designed a construct to express GFP and Sox10 under the control of the same promoter (pSSFV-*Sox10*-IRES-*GFP*) in order to detect the transduced cells (Panel A in [S1 Fig](#pone.0203785.s001){ref-type=\"supplementary-material\"}) and injected as viral particles into the corpora callosa of the mice after confirmation of demyelination using Y maze. ",
"In order to determine the identity of transduced cells, the pSFFV-IRES-*GFP* lentiviral particles which label the transduced cells without changing their fate, were injected into the corpora callosa (Panel B in [S1 Fig](#pone.0203785.s001){ref-type=\"supplementary-material\"}). ",
"After 21 days, the animals were sacrificed for brain sampling and IHF studies against different cell markers. ",
"Accordingly, most transduced cells tested positive for Gfap as a marker of astrocytes ([Fig 2A](#pone.0203785.g002){ref-type=\"fig\"}), while oligodendrocyte lineage cells comprised a small number of transduced cells ([Fig 2B--2D](#pone.0203785.g002){ref-type=\"fig\"}). ",
"There were no prominent co-expressions of NeuN (a neuronal marker) and GFP detected, which indicated that the neurons were not transduced. ",
"The lack of transduction appeared to be due to their distinct location from the injection site ([Fig 2E](#pone.0203785.g002){ref-type=\"fig\"}). ",
"Following quantification, the proportion of different cell types within the transduced GFP+ cells were as follows: 72.75%±4.3 (Gfap+), 13.5%±1.8 (PdgfRα+), 15%±2.8 (NG2+), and 11.25%±1.3 (Olig2+). ",
"No NeuN+/ GFP+ cells were detected ([Fig 2F](#pone.0203785.g002){ref-type=\"fig\"}). ",
"A part of above mentioned cells may be originated from the transduced progenitor cells which have been differentiated to astrocytes or oligodendroglial cells during 3 weeks chase time. ",
"Therefore the origin of transduced cells in this experiment remains not fully cleared.",
"\n\n{#pone.0203785.g002}\n\nWe investigated whether *Sox10* could convert astrocytes into oligodendrocyte lineage cells *in vivo*. ",
"Viral particles that contained the pSFFV-*Sox10*-IRES-*GFP* (*Sox10*-*GFP*) vector were directly delivered into the corpora callosa of the cuprizone induced demyelinated mice, where reactive astrocytes persisted. ",
"Mice were followed for 3 weeks after the injection. ",
"Ultimately, the brain samples were collected and prepared for sectioning and staining against astrocyte marker Gfap ([Fig 3A](#pone.0203785.g003){ref-type=\"fig\"}), and oligodendrocyte markers PdgfRα ([Fig 3B](#pone.0203785.g003){ref-type=\"fig\"}), NG2 ([Fig 3C](#pone.0203785.g003){ref-type=\"fig\"}), and Olig2 ([Fig 3D](#pone.0203785.g003){ref-type=\"fig\"}). ",
"Interestingly, as the representative micrographs showed, higher numbers of GFP+/Olig2+, GFP+/NG2+, and GFP+/PdgfRα+ cells compared to GFP+/Gfap+ cells were counted. [",
"Fig 3E](#pone.0203785.g003){ref-type=\"fig\"} shows the quantified data for the percentages of different GFP+ cell types. ",
"Within the GFP+ cells, we observed significantly more Olig2+, NG2+, and PdgfRα+ cells compared to Gfap+ cells (all: p\\<0.001). ",
"When compared the percentage of GFP+ cells with different labels to those mentioned in [Fig 2](#pone.0203785.g002){ref-type=\"fig\"} (animals which received only-GFP expressing vector), there was a significantly reduced number for Gfap+ cells (72.75% vs. 20%). ",
"The number of GFP+ cells stained with oligodendrocyte lineage cell markers were significantly increased by Sox10 expression, e.g. NG2+ cells increased from 15% to 74% which purpose possible conversion of astrocytes to oligodendrocyte lineage cells.",
"\n\n{#pone.0203785.g003}\n\nIn another group of animals, *Sox10*-*GFP* expressing viral vectors were injected into the corpora callosa of demyelinated mice and the samples were collected 8 weeks after the injection. ",
"Staining against Mbp and Plp as markers of mature oligodendrocytes showed that transduced (GFP+) cells were mainly differentiated to oligodendrocytes-like cells ([Fig 4A and 4B](#pone.0203785.g004){ref-type=\"fig\"}). ",
"Data quantification showed that 70.66±2.6 and 62.25± 3.3 percent of GFP+ cells were respectively Mbp+ and Plp+ ([Fig 4C](#pone.0203785.g004){ref-type=\"fig\"}).",
"\n\n{#pone.0203785.g004}\n\nThe previous experiment showed that local injection of Sox10-expressing vector into the demyelinated corpus callosum increased the number of oligodendrocytes lineage-like cells within the transduced cells. ",
"Since the origin of in vivo transduced cells was not fully clear, it was not possible to make a solid conclusion that astrocytes had the capability to differentiate to oligodendrocyte-like cells after receiving the Sox10 expression vector. ",
"Therefore, we planned to extract primary astrocytes, purify and transduce them with Sox10 expressing vector for subsequent transplantation into demyelinated mice corpora callosa. ",
"To prepare the starting cells, we isolated astrocytes from the brains of 3--5 day-old mice ([S2 Fig](#pone.0203785.s002){ref-type=\"supplementary-material\"}). ",
"The primary culture cells underwent magnetic activated cell sorting against the CD44 cell surface marker to obtain a pure population of astrocytes. ",
"Astrocytes were negative for NG2, Sox10 and Olig2 as OPC and oligodendrocyte markers, and negative for Nestin as a marker for neural progenitors as evaluated by immunocytofluorescence ([S3 Fig](#pone.0203785.s003){ref-type=\"supplementary-material\"}). ",
"We transduced astrocytes *in vitro* and subsequently transplanted them into the corpora callosa of the cuprizone-induced demyelinated brains ([Fig 5A](#pone.0203785.g005){ref-type=\"fig\"}). ",
"After 3 weeks, the brain sections were stained for Gfap (astrocyte marker) and PdgfRα, NG2, Mbp, and Plp as markers of oligodendrocyte progenitors and mature oligodendrocytes. ",
"According to [Fig 5B--5F](#pone.0203785.g005){ref-type=\"fig\"}, GFP+ cells expressed PdgfRα and NG2, as markers of OPCs, as well as Mbp and Plp, as markers of myelinating oligodendrocytes. ",
"A low number of GFP+ cells were still Gfap+. ",
"As we observed in 6 independent animals, following transplantation, GFP only-transduced astrocytes could not survive at 3 weeks after transplantation. ",
"These data may provide an evidence that Sox10 but not GFP (as control) caused in vivo trans-differentiation of astrocytes to oligodendrocyte lineage cells which were capable to integrate into the tissue and survive.",
"\n\n{ref-type=\"fig\"}, astrocyte transduction, and transplantation. ",
"B) Immunohistofluorescence (IHF) staining against Gfap showed the lack of notable co-expressions of GFP and Gfap as astrocyte marker at 3 weeks post injection (wpi). ",
"C-D) Expressions of PdgfRα (C) and NG2 (D) markers in the main population of GFP+ cells in the corpus callosum at 3 wpi. ",
"The bottom panels show higher magnifications. ",
"E-F) IHF showed that transplanted astrocytes (GFP+) could express mature oligodendrocyte markers Mbp (E) and Plp (F) at 3 wpi. ",
"n = 3 animals, CysA: cyclosporine A for preventing rejection of transplanted cells, Scale bars: 50 μm.](pone.0203785.g005){#pone.0203785.g005}\n\nTo further support the process of in vivo reprograming of astrocytes by sox10, astrocytes were transduced in vitro by viral particles and subsequently cultured in OPC induction medium ([Fig 6A](#pone.0203785.g006){ref-type=\"fig\"}). ",
"Approximately during 16--21 days after induction, we observed morphological changes in GFP-Sox10 vector transduced cells but not in GFP transduced astrocytes ([Fig 6B--6D](#pone.0203785.g006){ref-type=\"fig\"}). ",
"Interestingly, in Sox10 treated cells, the large flat shape morphology shifted into bipolar spindle-like cells ([Fig 6C and 6D](#pone.0203785.g006){ref-type=\"fig\"}) with proliferation capacity in response to PDGF-AA, bFGF and SHH. ",
"The induced cells showed expression of exogenous Sox10 as well as enhanced expression of OPC markers including Olig2, Myrf and endogenous Sox10 as assessed using RT-PCR. ",
"These cells did not express Krox20 a peripheral neural cell marker which showed the lack of Schwan cell differentiation. ",
"Astrocytes transduced with GFP vector did not express oligodendrocytes markers (Panel A in [S4 Fig](#pone.0203785.s004){ref-type=\"supplementary-material\"}). ",
"The expressions of exogenous and endogenous Sox10 was measured using differentiating primers. ",
"Olig2 expression was increased during days 5--9 post induction (Panel B in [S4 Fig](#pone.0203785.s004){ref-type=\"supplementary-material\"}). ",
"Compared to GFP vector transduced astrocytes, induced OPC-like cells showed decreased levels of Gfap and S100b expression (Panel C in [S4 Fig](#pone.0203785.s004){ref-type=\"supplementary-material\"}).",
"\n\n{#pone.0203785.g006}\n\nExpressions of the OPC markers were confirmed at the protein level by ICF against NG2 and Olig2 ([Fig 6E and 6F](#pone.0203785.g006){ref-type=\"fig\"}). ",
"Induced cells were negative for Tuj1 expression as a marker of neurons ([Fig 6G](#pone.0203785.g006){ref-type=\"fig\"}). ",
"Quantification of positive cells showed that 88.33%±2.1 of GFP positive cells exposed to induction OPC medium were NG2+, 74.33%±1.2 were Olig2+ and no Tuj1+ cell was detected ([Fig 6H](#pone.0203785.g006){ref-type=\"fig\"}).",
"\n\nDiscussion {#sec011}\n==========\n\nSox10 is related to the SRY gene family that regulates myelin gene expression in embryonic and adult oligodendrogenesis \\[[@pone.0203785.ref036]\\]. ",
"This factor binds to the enhancer of genes important for myelination, such as Myrf \\[[@pone.0203785.ref037]\\], Olig2 \\[[@pone.0203785.ref027]\\], and Mbp \\[[@pone.0203785.ref038]\\], and promotes oligodendrocyte differentiation.",
"\n\nIn situ conversion of reactive astrocytes present in the glial scars to progenitor cells such as OPCs suggests a new attitude toward myelin repair by replacement of reduced oligodendrocytes. ",
"This strategy also enhances repair potential by reducing the inhibitory components of glial scars. ",
"The outcomes of the current study have shown that astrocytes could possibly be converted into oligodendrocyte lineage cells by ectopic expression of a single transcription factor, Sox10. ",
"This finding supported previous reports where Sox10 in accordance with two other transcription factors reprogrammed rodent fibroblasts into iOPCs \\[[@pone.0203785.ref025], [@pone.0203785.ref026]\\]. ",
"Here, we used one out of the three previously reported transcription factors. ",
"Astrocytes have been used as the starting cells because they share a common embryonic origin with oligodendrocytes and contain extensive similarities in their epigenomes. ",
"In this study, we choose a single gene. ",
"A report showed the effectiveness of Sox10 to convert satellite glia into oligodendrocyte-like cells in the peripheral nervous system \\[[@pone.0203785.ref027]\\]. ",
"Several studies have shown the plasticity of astrocytes to reprogram into iPSCs \\[[@pone.0203785.ref039], [@pone.0203785.ref040]\\], NSCs \\[[@pone.0203785.ref011]\\], neuroblasts \\[[@pone.0203785.ref012]\\], and mature neurons \\[[@pone.0203785.ref010]\\]\\[[@pone.0203785.ref014]\\]. ",
"The reactive astrocytes within active demyelinated lesions are hypertrophic and proliferative, with some neural stem cell-like properties \\[[@pone.0203785.ref006]\\] that make them prone for reprogramming to progenitors. ",
"In addition, it has been reported that Sox10 can suppress astrocytic differentiation of glial progenitors via antagonistic relationship with NFIA as a key astrocyte fate determinant factor \\[[@pone.0203785.ref028]\\]. ",
"This may explain why Sox10 positive cells extracted from early post-natal brains mainly differentiate to oligodendrocyte \\[[@pone.0203785.ref029]\\]. ",
"Therefore NFIA inhibition by Sox10 may provide a possible mechanism for conversion of transduced astrocytes to oligodendrocyte lineage cells.",
"\n\nHere we have attempted to show the possible potency of Sox10 for in situ conversion of reactive astrocytes to OPCs. ",
"We observed reactivation of astrocytes in the brains of mice treated with cuprizone for 12 weeks. ",
"The viral particles were delivered to the corpora callosa of mice in order to target the astrocytes. ",
"At 3 weeks after the injection, astrocytes comprised 72.75% of GFP+ cells that received GFP control vector which imply for the astrocyte fate of the major group of transduced cells. ",
"Of course it should be mentioned that some labeled astrocytes or oligodendroglial cells observed in this experiment might be originated from progenitor cells transduced by GFP expressing viral vector at the time of viral particles injection. ",
"Most GFP+ cells that received Sox10 expressing vector expressed OPC markers Olig2 (73.75%), PdgfRα (54%), and NG2 (74%). ",
"Therefore a shift from astrocytes to OPCs by Sox10 was possibly occurred. ",
"At 8 weeks after the injection, some of the transduced cells expressed Mbp and Plp, as markers of mature oligodendrocytes. ",
"While the expression of ectopic Sox10 contributed to the conversion of transduced cells to OPCs, the role of the internal microenvironment of the demyelinated lesion should not be overlooked. ",
"Our previous experiments showed that *in vivo* ectopic expression of MicroRNA 302/376 in astrocytes converted them to neuroblasts \\[[@pone.0203785.ref015]\\], whereas application of the same microRNA into the brains of cuprizone fed mice caused transdifferentiation of astrocytes to oligodendrocyte lineage cells \\[[@pone.0203785.ref017]\\] which indicate the determining role of microenvironment.",
"\n\nThe enhanced number of OPCs 3 weeks after injection of the viral particles might be due to the conversion of astrocytes to OPCs; but other possibilities including proliferation of transduced OPCs and death of transduced astrocytes, or possible transduction of neural progenitors and their subsequent differentiation to OPCs by Sox10 might be plausible. ",
"Therefore, in order to prove conversion of astrocytes to OPCs, we extracted and purified astrocytes from the cortexes of P3-P5 mice. ",
"After transduction with viral particles that expressed GFP or GFP/Sox10, we transplanted the cells to the demyelinated mice brains and traced the transplanted cells at 3 weeks after transplantation. ",
"Results showed that transplanted cells expressed oligodendrocyte lineage cell markers, but in low amounts the astrocyte marker. ",
"Because of the very local conversion of astrocytes to oligodendrocytes within the injection site it was not expected that the transdifferentiated cells improve the animal behavior in Y-maze, while myelin staining for transplanted cells was observed. ",
"To support the proof of concept for the conversion of astrocytes to an oligodendrocyte lineage by Sox10, we attempted to show the possible potency of Sox10 for in vitro conversion of cultured astrocytes to OPCs. ",
"Our findings demonstrated that ectopic expression of Sox10 in astrocytes led to the expression of endogenous Sox10, Myrf and Olig2 in vitro. ",
"Expression of these key transcription factors was accompanied with supporting morphological changes.",
"\n\nIn this study we used a continuous active promoter to express Sox10, however the endogenous Sox10 and Olig2 locus became active in induced OPC-like cells as shown in [S4 Fig](#pone.0203785.s004){ref-type=\"supplementary-material\"}; accordingly converted cells might be a stable lineage independent on the transgene. ",
"The controls, the GFP vector-transduced cells in the oligodendrocyte media, lacked the ability to transdifferentiate to OPC-like cells which showed a pivotal role for Sox10. ",
"Sox10 function highly depends on its expression level \\[[@pone.0203785.ref041]\\]. ",
"However, we were unable to manage the exact copy number of ectopic genes expressed in each cell. ",
"This might explain why a population of infected cells were not converted to oligodendrocyte lineage.",
"\n\nUltimately, our proof-of-principle study may suggest the achievability of in vivo astrocyte conversion into oligodendrocyte-like cells using Sox10 gene delivery into the demyelinated lesions. ",
"Our data are promising for finding a new approach for myelin repair and astrocyte removal in patients with demyelinating insults.",
"\n\nSupporting information {#sec012}\n======================\n\n###### Schematic diagram of the expression vectors used in this study as well as the experimental procedure.",
"\n\nA\\) The expression vector contained open reading frame sequences of mouse Sox10 and GFP under the control of a single promoter, SSFV. ",
"Control vector did not include Sox10 sequence. ",
"B) Schematic diagram of the experimental procedure for inducing demyelination, Y maze test for data presented in [Fig 1D and 1E](#pone.0203785.g001){ref-type=\"fig\"}, virus injection, and tissue sampling.",
"\n\n(TIF)\n\n###### \n\nClick here for additional data file.",
"\n\n###### The morphology and characterization of isolated astrocytes before sorting.",
"\n\nA\\) immunocytofluorescence (ICF) showed that isolated astrocytes were positive for the glial fibrillary acidic protein (Gfap). ",
"We observed few Nestin (B), NG2 (C) and Olig2 (D) positive cells in cultured primary astrocytes. ",
"Scale bars: 100 μm.",
"\n\n(TIF)\n\n###### \n\nClick here for additional data file.",
"\n\n###### Determining the purity of sorted astrocytes.",
"\n\nAfter sorting the astrocytes using anti-CD44 antibody, we checked the purity of sorted astrocytes by immunocytofluorescence (ICF). ",
"ICF showed that purified cells were positive for glial fibrillary acidic protein (Gfap) (A) and negative for neural stem cell marker, Nestin (B), oligodendrocyte lineage markers Olig2 (C), Sox10 (D) and NG2 (E). ",
"Scale bars: 50 μm.",
"\n\n(TIF)\n\n###### \n\nClick here for additional data file.",
"\n\n###### Induction of astrocytes to oligodendrocytes by exogenous Sox10 led to the expressions of endogenous OPC markers.",
"\n\nA\\) Agarose gel electrophoresis analysis at day 21 after transduction confirmed the expression of endogenous Sox10, Olig2 and Myrf in cells transduced with Sox10-GFP vector, but not in astrocytes transduced with GFP vectors. ",
"B) Olig2 expression was increased in induced cells during days 5--9. ",
"C) Expression of Gfap and S100b as markers of astrocytes were reduced in induced OPC-like cells. ",
"Data in B and C was obtained using RT-qPCR. ",
"Primer specifications are provided in [S2 Table](#pone.0203785.s006){ref-type=\"supplementary-material\"}.",
"\n\n(TIF)\n\n###### \n\nClick here for additional data file.",
"\n\n###### List of primary and secondary antibodies used in this study.",
"\n\n(DOCX)\n\n###### \n\nClick here for additional data file.",
"\n\n###### Primer sets used for RT-qPCR analysis.",
"\n\n(DOCX)\n\n###### \n\nClick here for additional data file.",
"\n\n[^1]: **Competing Interests:**The authors have declared that no competing interests exist.",
"\n"
] | {
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[
"Create a Diaper Cake Towel for the baby shower.",
"Use these baby diaper cake kit to complete the look. ",
"It is sure to be a hit with the mom-to-be and all the guests.",
"\nKit Includes: ABC Box, Checkered ribbons, baby feet cu-outs as shown.",
"\nTowels are not included."
] | {
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[
"\"Well?\" \"",
"The target is still in the same place.\" \"",
"Target is leaving the apartment.\" \"",
"Ten seconds.\" \"",
"Start the engine.\" \"",
"Underwear or no underwear today?\" \"",
"Be a little more professional, guys.\" \"",
"She's turning left.\" \"",
"She's going to her usual cafe.\" \"",
"May I take your order?\" \"",
"A tea with milk, an orange juice and...\" \"And a croissant.\" \"",
"It's already ordered.\" \"",
"Thank you.\" \"",
"Do you have a visual?\" \"",
"We have a visual.\" \"",
"She's drinking her tea.\" \"",
"Come again?\" \"",
"She's drinking her tea!\" \"",
"Did you ask for a tea, sir?\" \"",
"No, no.\" \"",
"Thank you.\" \"",
"I'm looking for Ms. Elise Ward.\" \"",
"That's Ms. Ward over there.\" \"",
"Elise Ward?\" \" ",
"Yes, it's me.\" \" ",
"I have an envelope for you.\" \"",
"Somebody is talking to her.\" \"",
"Looks like a courier.\" \" ",
"Yes?\" \" ",
"She's at her usual cafe.\" \"",
"She's speaking with a man.\" \"",
"Looks like a messenger, you said to call if anything changed in her routine.\" \"",
"Stay on his face.\" \"",
"His face.\" \"",
"Do you think it's Alexander Pearce?\" \"",
"I think everybody is Pearce.\" \"",
"No, no.\" \"",
"It's okay.\" \"",
"It's a gift.\" \"",
"Goodbye.\" \"",
"All right, grab him.\" \" ",
"Take him.\" \" ",
"Let's get him.\" \"",
"Sir, stop!\" \"",
"Police!\" \"",
"What's going on?\" \" ",
"You are under arrest.\" \" ",
"Financial Crimes Division.\" \"",
"Division what?\" \"",
"Guys, I'm working!\" \"",
"Elise, you have no reason to trust me anymore but give me a chance to explain myself.\" \"",
"I know the police are watching you.\" \"",
"We have to throw them off the trail.\" \"",
"Board the 8:22 at the Gare de Lyon pick someone my height and build and make them believe it is me.\" \"",
"Burn this letter.\" \"",
"It is important you follow my instructions precisely.\" \"",
"I love you.\" \"",
"Alexander.\" \"",
"It's from him.\" \"",
"Save that letter.\" \"",
"Go in now.\" \"",
"Save that letter.\" \"",
"Daniel, get over there now.\" \"",
"Well?\" \"",
"The letter is burnt to shit.\" \"",
"Don't fucking touch it.\" \"",
"Take the ashes, take them to the van.\" \"",
"I'll send you someone.\" \"",
"Drive around to the other side!\" \"",
"Go!\" \"",
"Turn right at the end.\" \"",
"Faster!\" \"",
"Wednesday meeting.\" \"",
"Specialist pool.\" \"",
"So Alexander Pearce has a new face just as you suspected.\" \"",
"Thought it must've taken a lot of the money he stole from Reginald Shaw to finance this transformation.\" \"",
"Achmed Tchebali.\" \"",
"Bicycle courier of Algerian descent.\" \"",
"Not only has Pearce acquired a wife and child he's also 4 inches shorter than he used to be.\" \"",
"That must have been a big item of expenditure.\" \"",
"Congratulations, Acheson.\" \"",
"You have, indeed, cracked this case.\" \"",
"Letter to Inspector Collins.\" \"",
"Sir, this morning, she received a note, we have strong reason to believe...\" \"Alexander Pearce has 744 million in illegal assets that, given he is a British subject, we might seize.\" \"",
"Fraud complaints.\" \"",
"This operation, so far, has cost me 8 million.\" \"",
"If I thought there were more than a one-in-a-hundred chance you could be successful it would be rational for me to continue this operation.\" \"",
"Money laundering.\" \"",
"I do not.\" \" ",
"Yes, that's an amendment.\" \" ",
"Yes.\" \" ",
"There.\" \" ",
"It needs redrafting.\" \"",
"Yes, sir.\" \" ",
"Everything after code seven.\" \" ",
"Thank you, sir.\" \"",
"There's just one more.\" \"",
"Home Office.\" \"",
"Yes, and that's it.\" \"",
"Yes.\" \"",
"That's it.\" \"\"",
"Eight twenty-two.\"\" \" ",
"Projection for the pension plan fraud?\" \" ",
"Not now.\" \"\"",
"Eight twenty-two.\" \"",
"Lyon.\"\" \"",
"Are they meeting in Lyon at 8:22?\" \" ",
"Gare de Lyon, perhaps?\" \" ",
"Gare de Lyon.\" \"",
"That's it.\" \"",
"Gare de Lyon.\" \"",
"The train station.\" \"",
"She's getting on an 8:22 train.\" \"",
"Go.\" \"\"",
"Board the 8:22 at the Gare de Lyon.\"\" \"\"",
"Pick someone my height and build, make them believe it is me.\"\" \" ",
"I'm sorry.\" \" ",
"What for?\" \"",
"Mind me smoking?\" \"",
"It's not a real cigarette.\" \" ",
"What?\" \" ",
"It's electronic.\" \"",
"Delivers the same amount of nicotine, but the smoke is water vapor.\" \"",
"Yeah.\" \"",
"Watch.\" \"",
"LED light.\" \"",
"That's somewhat disappointing.\" \"",
"Would you have me smoking for real?\" \"",
"I would rather you be a man who did exactly as he pleased.\" \" ",
"l'm Elise.\" \" ",
"l'm Frank.\" \"",
"That's a terrible name.\" \"",
"It's the only one I've got.\" \" ",
"Maybe we can find you another.\" \" ",
"Okay.\" \"",
"You're British.\" \"",
"I'm American.\" \"",
"What brings you to Venice?\" \"",
"You read spy novels.\" \"",
"I'm a mysterious woman on a train.\" \"",
"You tell me what my story is.\" \"",
"Okay.\" \"",
"I think you'd be a diplomatic attache.\" \"",
"Or maybe a girl from East Germany whose father's been kidnapped and they're blackmailing you into stealing something for them.\" \"",
"A microfilm.\" \"",
"There's usually microfilm involved.\" \" ",
"What awaits me?\" \" ",
"Trouble, certainly.\" \" ",
"Danger?\" \" ",
"Oh, yeah.\" \"",
"You'll be shot at in less than two chapters.\" \"",
"Is there a man in my life?\" \"",
"Have to wait and see.\" \" ",
"Invite me to dinner, Frank.\" \" ",
"What?\" \"",
"Would you like to have dinner?\" \"",
"Women don't like questions.\" \" ",
"Join me for dinner.\" \" ",
"Too demanding.\" \"",
"Join me for dinner?\" \"",
"Another question.\" \"",
"I'm having dinner if you'd care to join me.\" \"",
"Musician.\" \"",
"Bartender.\" \"",
"I give up.\" \"",
"Math.\" \"",
"I teach math.\" \"",
"I would not have guessed that.\" \"",
"I'd imagine you're the cool math teacher.\" \"",
"Still a math teacher.\" \"",
"How about a photo?\" \"",
"Go right ahead.\" \"",
"Make me look good.\" \"",
"Could this be Pearce?\" \"",
"We still working on that?\" \"",
"Well, I only have the sketches to go by, so....\" \"Ectomorph, yes.\" \"",
"Same basic phenotype.\" \" ",
"Bottom line, could be him.\" \" ",
"Knew he couldn't leave her.\" \"",
"Run a worldwide blind check on that.\" \"",
"I bet you 1 0 quid there's no match.\" \"",
"Okay, what case number do you want to book it under?\" \" ",
"Just run it.\" \" ",
"Do you really think it's Pearce?\" \"",
"Run it.\" \"",
"Get me Italian Interpol.\" \"",
"I have a strange feeling that those two people over there are watching us.\" \"",
"Do you know something?\" \"",
"I think you're right.\" \"",
"Really?\" \"",
"You're the paranoid math teacher.\" \"",
"Memorize his face.\" \"",
"The train arrives in ten minutes.\" \"",
"As soon as he steps off, arrest him.\" \"",
"So, what are you doing all alone in the city of lovers?\" \"",
"Is there no one in your life?\" \"",
"There was.\" \"",
"What happened?\" \"",
"She left me.\" \"",
"I'm sorry to hear that, Frank.\" \"",
"Arrival at train station Venezia Santa Lucia in 1 0 minutes.\" \"",
"End of the line.\" \"",
"Guess I should go get my suitcase and stuff.\" \"",
"Goodbye.\" \"",
"Thank you.\" \"",
"Oh, shit.\" \" ",
"Sir, I...\" \" Wait.\" \"\"",
"Pick someone my height and build and make them believe it's me.\"\" \"",
"We did get an exact match.\" \"",
"He's a math teacher from Madison Community College.\" \"",
"Frank Tupelo.\" \"",
"Lost his wife in a car crash three years ago.\" \"",
"He's a tourist.\" \"",
"Thank you.\" \"",
"Get me Italian Interpol.\" \"",
"Understood.\" \"",
"The operation has been canceled.\" \"",
"Return to headquarters.\" \"",
"Quickly.\" \"",
"On the other side.\" \"",
"What are you doing?\" \"",
"The Internet fraud report.\" \"",
"So this is what Alexander Pearce looks like.\" \"",
"That's above your clearance.\" \"",
"Get back to your work.\" \"",
"He's fluttering his eyelids like a girl.\" \"",
"That's his tell.\" \"",
"Go all in.\" \"",
"Yes?\" \"",
"I have a piece of information for Mr. Shaw.\" \"",
"It's concerning Alexander Pearce.\" \"",
"Alexander Pearce just arrived in Venice with her.\" \" ",
"Valeriy?\" \" ",
"Sir?\" \"",
"Change of course.\" \" ",
"Venice.\" \" ",
"Yes, sir.\" \"",
"Now, what do I own in Venice?\" \"",
"Frank!\" \"",
"Hi.\" \" ",
"Do you want to come with me?\" \" ",
"What?\" \"",
"Oh, yeah, sure.\" \"",
"I mean, I could use a lift, yeah.\" \"",
"Thank you very...\" \"That's very kind.\" \"",
"Welcome to Venezia, signora.\" \"",
"The bags.\" \"",
"Oh, my bags.\" \"",
"Are you coming?\" \"",
"Elise Clifton-Ward and husband.\" \"",
"Oh, we arranged for you a wonderful suite with a bathroom and with....\" \" May I have your documents?\" \" ",
"Yes.\" \"",
"Thank you.\" \"",
"It would be my pleasure to show you up personally.\" \" ",
"Please, this way.\" \" ",
"Thank you.\" \"",
"Enjoy your stay, sir.\" \"",
"Thank you.\" \"",
"Everything was arranged according to the precise instructions.\" \"",
"I very much hope you will not be disappointed.\" \"",
"We are entering the Doge's Suite.\" \"",
"It is a room that has been a home away from home for Marcel Proust and Honore de Balzac and others.\" \" ",
"Thank you.\" \" ",
"Oh, thank you.\" \"",
"Have a nice stay, sir.\" \"",
"Thank you.\" \"",
"Make yourself at home.\" \"",
"This really is a very nice hotel, isn't it?\" \"",
"They think of everything, don't they?\" \"",
"We know he will be with her.\" \"",
"They'll be staying at one of the hotels.\" \"",
"The Gritti, the Regina, the Danieli.\" \"",
"I want you to keep a watch on all of them.\" \"",
"You can kill the girl, but you can't kill him.\" \"",
"Not until I have my money, that is.\" \"",
"Understand?\" \"",
"Let's go.\" \"",
"Fuck.\" \" ",
"You're ravenous.\" \" ",
"Do you mean ravishing?\" \"",
"I do.\" \"",
"You're ravenous.\" \"",
"I am.\" \"",
"Come on.\" \"",
"The scampi and champagne risotto, please.\" \"",
"Excellent choice.\" \"",
"Excellent.\" \"",
"He didn't say that to me.\" \"",
"Did he not like my choice?\" \"",
"Your choice was excellent.\" \"",
"Who is he?\" \"",
"Hotels don't leave gifts like that, do they?\" \"",
"Who is he?\" \"",
"He's someone who used to be very important to me.\" \"",
"Used to?\" \"",
"Are you meeting him in Venice?\" \"",
"I hope so.\" \"",
"Part of me hopes I don't.\" \"",
"Why?\" \"",
"Because I don't like being told what to do.\" \"",
"I don't like being summoned.\" \"",
"But it felt even worse not hearing from him for two years.\" \"",
"It's the Roman god, Janus.\" \"",
"My mother gave it to me when I was little.\" \"",
"She wanted it to teach me that people have two sides.\" \"",
"A good side, a bad side.\" \"",
"A past, a future.\" \"",
"And that we must embrace both in someone we love.\" \"",
"And I tried.\" \"",
"What's he like?\" \"",
"He's different from anybody I know.\" \"",
"Different's good.\" \"",
"Where I come from the highest compliment they can offer a person is to say that they're down-to-earth.\" \"",
"Grounded.\" \"",
"I hate it.\" \"",
"It drives me nuts.\" \"",
"Would you like another Americano?\" \"",
"I don't know.\" \"",
"Would I?\" \"",
"That really was a very nice restaurant, wasn't it?\" \"",
"A very nice wine.\" \"",
"I'm making us a nice little drink which is very nice of me, isn't it?\" \"",
"Thank you.\" \"",
"Were those flowers here when we checked in?\" \"",
"It's an invitation to a ball two nights from now.\" \"",
"You've been summoned.\" \"",
"It appears I have.\" \"",
"May I pay you a compliment?\" \"",
"Another question you need never ask.\" \"",
"You are the least down-to-earth person I've ever met.\" \"",
"It's been a long day.\" \"",
"I do hope the couch will be comfortable.\" \"",
"Good night.\" \"",
"Good morning.\" \"",
"My name is Guido.\" \"",
"I'm your waiter.\" \"",
"I'm Frank.\" \"",
"What's this?\" \"",
"It's your breakfast, sir.\" \"",
"I didn't order any.\" \"",
"La signora order it for you when she left.\" \"",
"When she left?\" \"",
"Have a nice day, sir.\" \" ",
"Hi.\" \" ",
"Alexander.\" \"",
"Let's go see an old friend.\" \"",
"What?\" \" ",
"He doesn't look like Pearce.\" \" ",
"Boss said he might not.\" \"",
"Grab him!\" \"",
"Hello?\" \"",
"Please help.\" \"",
"Bon Jovi.\" \"",
"I need help.\" \"",
"There are two men trying to break down the door.\" \"",
"I see.\" \"",
"What problem do you have with the door?\" \"",
"Two men with guns trying to break in.\" \" ",
"Your Spanish is excellent, sir.\" \" ",
"Thank you.\" \" ",
"l gotta go!\" \" ",
"Sir?\" \"",
"Americans....\" \"Boss wants him alive, you idiot!\" \"",
"She's coming back.\" \"",
"Frank!\" \"",
"Frank.\" \"",
"Prepare to intervene.\" \" ",
"Yes.\" \" ",
"Commander Acheson two armed men are chasing the American.\" \"",
"Shots have been fired.\" \" ",
"Request permission to intervene.\" \" ",
"Do you still have Elise?\" \"",
"We still have a visual but the American is in imminent danger.\" \"",
"The mathematics teacher from Wisconsin?\" \"",
"I'd say not our mandate.\" \"",
"Sir, this man is in real peril.\" \"",
"This is exactly why she chose him.\" \"",
"To distract us.\" \"",
"Stay with her.\" \"",
"Do not move unless she moves.\" \"",
"Am I making myself crystal clear?\" \"",
"Sir, they will kill him.\" \"",
"Not our mandate.\" \"",
"Hello?\" \"",
"Hello?\" \"",
"Hello?\" \"",
"Oh, God.\" \"",
"Headquarters!\" \"",
"Send me backup to the fruit market.\" \"",
"I have a nutcase who wants to jump.\" \"",
"Probably American.\" \"",
"Stop, stop!\" \"",
"Please!\" \"",
"Don't do it!\" \"",
"Oh, my God.\" \"",
"I'm so sorry.\" \"",
"Bastard!\" \"",
"I'm so sorry.\" \"",
"Stop him!\" \"",
"I think you need it more than I.\" \"Thank you.\" \"",
"You smoke?\" \"",
"No.\" \"",
"Now.\" \"",
"You wish to report a murder.\" \"",
"No.\" \"",
"Some people tried to kill me.\" \"",
"I was told you were reporting a murder.\" \"",
"Attempted murder.\" \"",
"That's not so serious.\" \"",
"No.\" \"",
"Not when you downgrade it from murder.\" \"",
"But when you upgrade it from room service, it's serious.\" \"",
"So is assaulting a police officer.\" \" ",
"That was an accident.\" \" ",
"The officer feels differently.\" \"",
"With all due respect, sir, I think maybe I should be talking with someone...\" \"Someone not ltalian.\" \"",
"Like the embassy or Interpol.\" \"",
"Interpol.\" \"",
"I'm an American citizen.\" \" ",
"What does that mean?\" \" ",
"l'm involved in something here and no offense, this is not a local issue.\" \"",
"What is it you think you're involved in?\" \"",
"It all started when I met a woman on the train from Paris.\" \" ",
"This is already good.\" \" ",
"No.\" \"",
"No.\" \"",
"She took me to her hotel.\" \"",
"I don't know.\" \"",
"No.\" \"",
"She's involved with another man.\" \"",
"She's in love.\" \"",
"I'm sorry to hear that.\" \"",
"I know.\" \"",
"Anyway, this man, whatever his story is some people are trying to kill him.\" \"",
"How do you know?\" \"",
"They tried to kill me.\" \"",
"They tried to kill you?\" \"",
"Yes.\" \"",
"Because they must have thought that I was him.\" \"",
"I like you, Mr. Tupelo.\" \"",
"I like you too, sir.\" \"",
"But surely you understand how strange this all sounds?\" \"",
"Come with me.\" \"",
"Where are you taking me?\" \" ",
"Oh, you're gonna lock me in there?\" \" ",
"Don't worry.\" \"",
"Please don't do this.\" \"",
"Sir, I've done nothing wrong.\" \" ",
"Please.\" \" ",
"lt's only until I check some facts.\" \"",
"Hi.\" \"",
"Your facts checked out.\" \"",
"We have to leave.\" \"",
"You're not safe here.\" \"",
"I don't understand.\" \"",
"This has to look right.\" \"",
"Come on.\" \"",
"The man you are talking about is called Alexander Pearce.\" \"",
"He stole big money from a gangster.\" \"",
"They've come to Venice to find him.\" \"",
"They think you are him.\" \"",
"They've placed a....\" \"A bounty on your head.\" \"",
"That's why you're not safe.\" \"",
"Wait here for a moment.\" \"",
"You've come to buy a key?\" \"",
"Brand-new bills?\" \"",
"I appreciate it.\" \" ",
"You think we cheat you?\" \" ",
"Of course not.\" \"",
"I just enjoy counting.\" \"",
"Well, gentlemen, Alexander Pearce is all yours.\" \"",
"Remember, don't shoot him!\" \"",
"I think we've lost them.\" \"",
"I'd really like to take these off.\" \"",
"We have to wait to get to open waters.\" \"",
"Okay.\" \"",
"Elise!\" \"",
"Elise!\" \"",
"Elise!\" \"",
"You all right?\" \"",
"Yeah.\" \"",
"Do you think it's a little loose?\" \"",
"No, no, no.\" \"",
"They are back.\" \"",
"Bring them in.\" \"",
"I come back tomorrow?\" \"",
"No, no.\" \"",
"No, stay.\" \"",
"We tried everything, sir.\" \" ",
"But he got away.\" \" ",
"So I heard.\" \"",
"So I heard.\" \"",
"They're referring to the young man who stole $2.3 billion from me.\" \"",
"You know, I worked very hard for years so I wouldn't have to do the dirty work.\" \"",
"No more chasing traitors.\" \"",
"No more killing.\" \"",
"Now I employ people to do those things for me.\" \"",
"Except that you don't.\" \" ",
"Does it look all right?\" \" ",
"Yes.\" \"",
"Like always.\" \"",
"What's \"always\"?\" \"",
"Very good.\" \"",
"Yes, I think it looks all right.\" \"",
"So that's how you pick a lock.\" \"",
"Why do you know how to do that?\" \"",
"Where's the owner of this boat?\" \"",
"Bottom of a canal?\" \"",
"Sitting in a cafe with a pocketful of euros.\" \"",
"Why is all this happening?\" \"",
"Why is everyone trying to kill me?\" \"",
"It's because I kissed you.\" \"",
"Do I look that much like him?\" \"",
"I am sorry l got you involved in all this.\" \"",
"Why are you involved in all this?\" \" ",
"Because I'm in love with him.\" \" ",
"Really?\" \"",
"I'm not.\" \"",
"Seriously, help me understand why.\" \"",
"I honestly didn't think he would let it go far.\" \"",
"I didn't think he would let Shaw get to you.\" \"",
"Shaw.\" \"",
"That's the man that Pearce stole from?\" \"",
"Reginald Shaw.\" \"",
"Alexander used to be his private banker.\" \"",
"I thought he'd be Russian or something like that.\" \"",
"He surrounds himself with Russians, but he's as English as Alex and I.\" \"Give me your hand.\" \"",
"Now he owns most of the casinos and brothels from here to Novosibirsk.\" \"",
"Once, he bragged to Alexander that he had every man killed his wife had slept with before she met him.\" \"",
"When he found out how many there'd been, he killed her too.\" \"",
"There you are.\" \"",
"What made Pearce think that he could take on a guy like that?\" \"",
"It's just who he is.\" \"",
"Could you ever feel like that about someone like me?\" \"",
"I don't regret it, you know.\" \"",
"Kissing you.\" \"",
"I have to go check the thing.\" \"",
"Where are we going?\" \"",
"The airport.\" \" ",
"Where are we going?\" \" ",
"You'll see.\" \"",
"Tie us off, would you?\" \"",
"I wish we'd met in another life, Frank.\" \"",
"Take the next flight to America.\" \"",
"Find what you need in the bag.\" \"",
"Be safe.\" \"",
"But I'm in love with you.\" \"",
"Agent Elise Clifton-Ward identification number MFS98495G.\" \"Elise Ward.\" \"",
"Well, you've certainly blown your cover now.\" \"",
"I'm ready to give you Pearce.\" \"",
"You're ready to give me Pearce.\" \"",
"That's interesting.\" \"",
"So you aren't here to beg me to lift your suspension.\" \"",
"You've actually come to do your job.\" \"",
"Allow me to ask why now?\" \"",
"You live with a man for a year during which time we don't get one single usable photograph.\" \"",
"But now you're ready to give him up.\" \"",
"It's because of that tourist, isn't it?\" \"",
"I want to put an end to this before someone gets killed.\" \"",
"I wish I could understand your choice in men.\" \"",
"A fugitive thief.\" \"",
"Well, that's exciting, perhaps.\" \"",
"But a schoolteacher from Wisconsin?\" \"",
"Who'd have thought your tastes were so provincial?\" \"",
"Do you want Pearce or not?\" \"",
"Yes.\" \"",
"I'm meeting him at a ball tonight.\" \"",
"You know you'll have to wear a wire.\" \"",
"Good.\" \"",
"She's heading toward the southeast corner.\" \"",
"Where have you been?\" \"",
"I thought you'd never get here.\" \"",
"Really?\" \"",
"Certainly.\" \"",
"Is that our man?\" \"",
"How could you be so sure I was coming?\" \"",
"I just knew it.\" \"",
"Fate wouldn't bring me to an evening like this with no reason.\" \"",
"As soon as you walked in, I knew what the reason was.\" \"",
"No?\" \"",
"No.\" \"",
"Count Filippo Gaggia.\" \"",
"Landowner.\" \"",
"Big reputation as a swordsman.\" \"",
"Well, he won't be dueling tonight.\" \"",
"Wait.\" \"",
"Alexander.\" \" ",
"Excuse me.\" \" ",
"lt's Pearce.\" \"",
"He's here.\" \"",
"He just left her that envelope.\" \"",
"Pull up the footage, we have his face.\" \"",
"Alexander.\" \"",
"Frank.\" \"",
"I don't bloody believe this.\" \"",
"All right, get that moron out of here.\" \"",
"You shouldn't...\" \"I can't, I'm sorry.\" \"",
"Come on.\" \"",
"You owe me a dance.\" \"",
"You shouldn't be here.\" \"",
"No, Elise, this is the one place on earth I should be.\" \"",
"How did you get in here?\" \"",
"I told the doorman you were my wife and I wanted to keep an eye on you.\" \"",
"Italians are big on that kind of thing.\" \" ",
"Leave.\" \" ",
"No.\" \"",
"Do you like the suit?\" \"",
"You paid for it.\" \"",
"I've been thinking about your friend Pearce and his plan.\" \"",
"And so far, I'm thinking it hasn't worked out for him.\" \" ",
"No?\" \" ",
"No.\" \"",
"Take that gangster guy.\" \" ",
"Shaw.\" \" ",
"Yeah, Shaw.\" \"",
"I don't believe Pearce was prepared for him being here for the whole chase.\" \"",
"Will you please leave?\" \"",
"You're kind of worried about me?\" \"",
"Yes.\" \"",
"And I'm worried about you.\" \"",
"Frank.\" \"",
"And I'm not leaving without you.\" \"",
"You were part of a plan.\" \"",
"You were useful, that's all.\" \"",
"What do you think, I saw you on the train and my heart stopped?\" \"",
"Leave.\" \"",
"This way, please.\" \"",
"Excuse me, sir, I think you've made a mista...\" \"What's it say?\" \"",
"Bring up Signora Ward's boat, please.\" \"",
"It looks like they've called up her boat.\" \"",
"What should we do?\" \"",
"She may just be playing with us.\" \"",
"Let's wait.\" \"",
"Fondamenta San Giacomo 23, please.\" \"",
"Okay, good.\" \"",
"We have the address.\" \"",
"I'm gonna follow her on the boat.\" \"",
"I want snipers on all surrounding roofs and if you get a visual on Pearce first, call.\" \"",
"Help.\" \"",
"Hello?\" \"",
"Let's have a little fun with him.\" \"",
"Who are you people?\" \"",
"Where are we going?\" \"",
"Well, I don't know, Alexander.\" \"",
"Where are we going?\" \"",
"I'm not Alexander.\" \"",
"My name is Frank Tupelo.\" \"",
"I'm an American citizen and I have rights.\" \"",
"I don't care what you call yourself these days.\" \"",
"Your name is Alexander Pearce, and you have no rights.\" \"",
"Not till you pay us 744 million pounds back taxes.\" \"",
"Seven hundred and forty-four million?\" \"",
"Looks like someone else is following our target.\" \"",
"Reginald Shaw.\" \"",
"Well, this could be fun.\" \"",
"Stay close but not too close, and get the office to send me an interpreter.\" \"",
"Yes, 744 million.\" \"",
"You didn't think we knew about that bank account in Liechtenstein.\" \" ",
"l don't know what he's talking about.\" \" ",
"Listen you asshole.\" \"",
"I know everything about you.\" \"",
"I even know how much that face cost.\" \"",
"Twenty-four million wired to a surgeon from a Swiss bank account.\" \"",
"I haven't been able to prove it but now I don't have to.\" \"",
"Because I just lock you in a box in a basement until you tell me where the money is.\" \"",
"It's all right.\" \"",
"It's all right, Frank.\" \"",
"I know you're not Alexander Pearce.\" \"",
"But you know who you are?\" \"",
"You're a moron.\" \"",
"And you're a moron that almost ruined my 8-million-pound sting operation.\" \"",
"Twice.\" \"",
"So you're gonna stay here until this is over.\" \"",
"Please, you can't do this.\" \"",
"I haven't done anything.\" \"",
"I shouldn't be here.\" \"",
"You know what I think?\" \"",
"I think that this is the one place on earth you really should be.\" \"",
"How was your night in the hotel, huh?\" \"",
"She's quite something, I'm sure.\" \"",
"Target is about 200 yards away from destination.\" \" ",
"We are matching speed.\" \" ",
"Good.\" \"",
"But don't get too close.\" \"",
"I don't wanna scare Pearce off again.\" \"",
"I won't be needing you anymore this evening.\" \"",
"Magnificent.\" \"",
"I've never really understood the love that people devote to each other.\" \"",
"Money, yes.\" \"",
"Possessions, yes.\" \"",
"But people are so unreliable.\" \"",
"They sicken, decay and die.\" \"",
"But this....\" \"This is worth dying for.\" \"",
"Alexander had taste.\" \" ",
"It seems to me that....\" \" We're live.\" \"",
"Oh, yes, do sit down.\" \"",
"When are you going in?\" \"",
"You're welcome to stay and watch, if you can keep your mouth shut.\" \"",
"You know, Alexander was like a son to me.\" \"",
"I loved him.\" \"",
"That brilliant, playful, disrespectful young bastard.\" \"",
"I even thought he and I were two of a kind.\" \"",
"Oh, I took great pleasure never knowing what the scamp would do next.\" \"",
"Until one day, what he did next didn't give me any pleasure at all.\" \"",
"You know, in our quaint legal system, if a man sleeps with my wife I kill him and her and get away scot-free.\" \"",
"Crime of passion is what it's called.\" \"",
"But my passion extends to all the things I own.\" \"",
"They are me.\" \"",
"They represent me.\" \"",
"So if a man steals from me, I kill him.\" \"",
"I kill his wife, his children, his mother.\" \"",
"I might even throw in the family doctor.\" \"",
"For he has taken from me something for which I have paid the infinite price:\" \"my soul.\" \"",
"That's something these idiots will never understand.\" \"",
"He's not coming, is he?\" \"",
"He sent you to get the money.\" \"",
"So where is it?\" \"",
"Where's my money?\" \"",
"Where is the safe?\" \"",
"I don't know.\" \"",
"Where is the safe?\" \" ",
"l don't know.\" \" ",
"You don't know?\" \"",
"Is it behind this bookcase here?\" \"",
"Or perhaps this one here.\" \" ",
"Where is the safe?\" \" ",
"l don't know.\" \"",
"I haven't seen Alex...\" \"Please, do something.\" \" ",
"Permission to fire.\" \" ",
"Hold your fire.\" \"",
"I repeat, hold your fire.\" \"",
"Not until Pearce shows.\" \"",
"You're very beautiful, Elise.\" \"",
"Now, try to combine it with a little intelligence and tell me where the safe is.\" \"",
"If I knew it, I would tell you.\" \"",
"I don't think she actually knows.\" \"",
"You have to go in.\" \"",
"They're gonna kill her.\" \" ",
"Where's my interpreter?\" \" ",
"They're going to kill her.\" \"",
"She can handle herself.\" \"",
"She can handle herself?\" \"",
"She's one of us.\" \"",
"Expert in undercover operations.\" \"",
"But when I sent her to Russia to find Pearce, she went dark.\" \"",
"I think she could never quite decide if she was with him or with us.\" \"",
"She's a good agent.\" \"",
"Except she falls in love with any man that she spends longer than a train ride with.\" \"",
"Commander Acheson, I'm the interpreter.\" \"",
"Yes, go.\" \"",
"If you could provide me with some context.\" \"\"",
"She really doesn't seem to know where it is.\" \"",
"We should go.\"\" \"\"",
"We give it five more minutes.\" \" ",
"lf Pearce doesn't show...\"\" \" Pearce.\" \"\"",
"If Pearce doesn't show, we get rid of the woman and go.\"\" \"",
"Permission to engage.\" \" ",
"Negative.\" \" ",
"Please.\" \"",
"Not one more word from you.\" \"",
"They won't kill her.\" \"",
"Pearce will come.\" \"",
"He has to come.\" \"",
"All right, let's do it.\" \"",
"No.\" \"",
"Knife.\" \"",
"Engaging target.\" \"",
"Negative.\" \"",
"That is an order.\" \"",
"This guy is crazy.\" \"",
"Now, if you don't tell me where the safe is I might be tempted to rearrange your face somewhat.\" \"",
"You'll find life is not quite so giving to an ugly woman.\" \"",
"You really don't know, do you?\" \"",
"There.\" \"",
"Behind the medallion.\" \"",
"There.\" \"",
"Open it.\" \"",
"There is movement in the courtyard.\" \"",
"Pearce. ",
"I knew it.\" \"",
"You know what?\" \"",
"He does look a little like you.\" \"",
"No!\" \" ",
"l don't...\" \" Just try.\" \"",
"Go on.\" \"",
"Press enter.\" \"",
"You may try one more time.\" \"",
"Let her go.\" \"",
"Oh, my God.\" \" ",
"You let her leave, I'll open the safe.\" \" ",
"Stop.\" \"",
"Frank, what in hell?\" \"",
"Please, Elise.\" \"",
"Let me handle this.\" \"",
"I'll say it again.\" \"",
"You let her go, I'll open the safe.\" \"",
"Who the hell are you?\" \"",
"I'm Alexander Pearce.\" \"",
"This was the man on the boat.\" \" ",
"He is not Alexander Pearce.\" \" ",
"Elise, please.\" \" ",
"What the hell are you doing?\" \" ",
"What the hell is he doing?\" \"",
"Permission to engage target.\" \"",
"Shut the hell up.\" \"",
"Alexander.\" \"",
"Is it you?\" \"",
"Truly?\" \"",
"Truly.\" \"",
"Show me your teeth.\" \"",
"Alexander had crooked teeth.\" \"",
"Stained from excessive smoking.\" \"",
"Braces.\" \"",
"Electronic cigarettes.\" \"",
"He had a higher forehead.\" \"",
"I had a migraine for two weeks after the remodel.\" \"",
"You sound different.\" \" ",
"Voice-chip implant.\" \" ",
"No, no.\" \"",
"I mean, the way you speak.\" \"",
"Yes, that's right.\" \"",
"Alexander was English.\" \"",
"This man is American.\" \"",
"I've gotten used to the American accent that I find it a tad difficult to go back.\" \"",
"That's pathetic.\" \"",
"This man is not Alexander Pearce.\" \"",
"He's a tourist.\" \"",
"A tourist?\" \"",
"Someone I happened to sit next to on a train.\" \"",
"How would a tourist know you killed every man your wife ever slept with?\" \"",
"I told him that.\" \"",
"You told a tourist?\" \"",
"Why would she do that?\" \"",
"It's absurd.\" \"",
"Well, there's a simple way of finding out.\" \"",
"Open the safe.\" \"",
"I've made you a deal.\" \" ",
"When she's gone...\" \" What are you doing?\" \"...",
"and I know she's safe, then....\" \"Well, I'll make you an even better deal.\" \"",
"Open the safe now, and I won't make you watch something that you would never forget.\" \"",
"Open it.\" \"",
"I see.\" \"",
"Now you see.\" \" ",
"Sir.\" \" ",
"l know him.\" \"",
"He'll come for her.\" \"",
"I know him.\" \"",
"He won't give her up now.\" \"",
"We must engage.\" \"",
"Permission to engage, sir.\" \"",
"Sir.\" \"",
"I love you.\" \"",
"He'll come for her.\" \"",
"He'll come for her.\" \"",
"It doesn't make se...\" \"This is Chief Inspector Jones.\" \"",
"Permission granted.\" \"",
"Fire.\" \" ",
"Are you all right?\" \" ",
"Yes.\" \"",
"Agent Ward, your suspension's lifted.\" \"",
"Thank you, sir.\" \"",
"And your employment is terminated.\" \"",
"Effective immediately.\" \"",
"Thank you, sir.\" \"",
"We got him.\" \" ",
"We got Alexander Pearce.\" \" ",
"Where?\" \"",
"On the Fondamenta.\" \"",
"Three hundred meters south of your location.\" \"",
"So you love me.\" \"",
"I do.\" \"",
"How do you feel about Alexander Pearce these days?\" \"",
"I love him too.\" \"",
"Unfortunately.\" \"",
"I may have a solution for you.\" \"",
"Are you with me this time?\" \"",
"Come on.\" \"",
"Let him up.\" \"",
"I'm just a tourist.\" \"",
"He said I might get arrested at some point.\" \" ",
"What?\" \" ",
"But that you'd have to let me go because you'd have nothing on me.\" \" ",
"Who?\" \" ",
"The man that's sending me texts.\" \" ",
"Texts?\" \" ",
"And the money, of course.\" \"",
"So let me get this straight because I'm a little confused.\" \"",
"You receive money from a man you've never met before who sends you text messages telling you just to show up somewhere.\" \"",
"Well not just anywhere.\" \"",
"Clear.\" \"\"",
"Seven hundred and forty-four million.\"\" \"",
"Inspector Acheson this operation is now officially terminated.\" \" ",
"l'll expect your report on Monday.\" \" ",
"Sir, listen.\" \" ",
"We have our money, Acheson.\" \" ",
"But we don't have Pearce.\" \"",
"The man's a criminal wanted in 14 countries.\" \"",
"Well, what is it he did, really?\" \"",
"He stole money from a gangster.\" \"",
"A dead gangster.\" \"",
"And he has good taste in women.\" \"",
"I can't say I don't wish him well.\" \"",
"Twenty million dollars' worth of plastic surgery.\" \"",
"And that's the face you choose?\" \"",
"Do you not like it?\" \"",
"It'll do.\""
] | {
"pile_set_name": "OpenSubtitles"
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[
"require 'unit_spec_helper'\n\ndescribe Rpush, 'embed' do\n before do\n allow(Rpush::Daemon).to receive_messages(start: nil, shutdown: nil)\n allow(Kernel).to receive(:at_exit)\n end\n\n after { Rpush.shutdown }\n\n it 'sets the embedded config option to true' do\n Rpush.embed\n expect(Rpush.config.embedded).to eq(true)\n end\n\n it 'starts the daemon' do\n expect(Rpush::Daemon).to receive(:start)\n Rpush.embed\n end\nend\n\ndescribe Rpush, 'shutdown' do\n before { Rpush.config.embedded = true }\n\n it 'shuts down the daemon' do\n expect(Rpush::Daemon).to receive(:shutdown)\n Rpush.shutdown\n end\nend\n\ndescribe Rpush, 'sync' do\n before { Rpush.config.embedded = true }\n\n it 'syncs' do\n expect(Rpush::Daemon::Synchronizer).to receive(:sync)\n Rpush.sync\n end\nend\n\ndescribe Rpush, 'status' do\n before { Rpush.config.embedded = true }\n\n it 'returns the AppRunner status' do\n expect(Rpush::Daemon::AppRunner).to receive_messages(status: { status: true })\n expect(Rpush.status).to eq(status: true)\n end\nend\n"
] | {
"pile_set_name": "Github"
} | [
0.003875968992248062
] | 0.003876 | 5 |
[
"Case 3. ",
"Localizing Chinese games into Russian\n\nDescription:\n\nA Chinese online game publisher is looking to localize a game. ",
"The deadlines are tight, the company has three contractors, the source content was written in English by Chinese speakers, there’s no time for testing, and there are no reference texts in Chinese. ",
"Prioritizing the deadline, the translations are handed to three different suppliers, with employees on staff tapped to handle the editing. ",
"Only one company suggests using a single software tool to ensure consistency and naming one of the three contractors responsible for complying with existing terminology and managing translation memories. ",
"For a variety of reasons, including the lack of time, the decision is made to just go with the existing options.",
"\n\nResult:\n\nAll three companies get the job done on time, though the quality level is poor. ",
"And since editing and compiling the texts needs to happen yesterday, there’s no time to figure out which companies performed better or worse. ",
"Only one of them gets any feedback. ",
"Which, you ask? ",
"The one that asked for it, of course. ",
"And from what that contractor can tell, the problems that were pointed out are all in sections of the translation handled by the other two.",
"\n\nBuilding processes within the localization department\n\nBesides setting goals, it’s important to keep a tight grip on translation quality. ",
"The best way to do that, in addition to randomization, is to institute a triple-blind system: the people being checked don’t know which part of their work will be checked, the testers don’t know whose work they’re checking, and the specialist processing the results doesn’t know who was checking who. ",
"Localization or linguistic testing is the simplest way to find and eliminate errors in the real world.",
"\n\nThere are two rules for making sure evaluations are objective:\n\nThe translation and testing teams are equidistant from the developer — if the translation is done by an external contractor, the testing should be as well. ",
"The translation and testing teams should be separate units with their own independent processes and staff. ",
"Translators shouldn’t moonlight as testers, and vice versa. ",
"That goes for both salaried and external teams.",
"\n\nLinguistic testing is a critical element of the localization process. ",
"Even experienced game translators sometimes don’t get the context or haven’t read the supporting documentation. ",
"Alternatively, a Chinese font might have been used for kanji in the game engine instead of a Japanese font, or tags might have been just stuck in due to a software quirk. ",
"Whatever the case, the outcome can be problematic. ",
"And that’s easier to catch when you’re playing through the game, making it important to have testers in place rather than dumping the issues on your players.",
"\n\nWhen it comes to a well-oiled localization process, the manager working for the developer needs a contractor to handle game translation and testing. ",
"That’s generally two separate companies, though there are some cases where a single supplier with mutually independent teams can take on both sides.",
"\n\nIf you have multiple contractors localizing a single project, it’s going to be almost impossible to maintain a uniform style throughout. ",
"There are going to be different kinds and quantities of issues in the resulting texts as well.",
"\n\nThe same is true of cases where you’re working with a single contractor, only they have more than three translators on the same project. ",
"That forces you to hire two or more editors who will have different linguistic styles and mistake profiles.",
"\n\nJust like in Brooks’ The Mythical Man-Month, linear expansion in the number of people working on a problem doesn’t exactly net you a similarly linear reduction in the project time frame. ",
"It can even start pushing deadlines back once you hit a certain number.",
"\n\nBut what do you do when deadlines are tight? ",
"That’s when it comes in handy to have a single glossary and a general translation memory that have to be steadily updated. ",
"And while either the developer’s localization manager or one of the contractors can take responsibility for doing that, it does need to be just one person. ",
"That will cut down on errors, though it won’t eliminate them.",
"\n\nAnd so the perfect translation is handled by one translator and one editor working within reasonable deadlines. ",
"That’s about 10,000 words a week.",
"\n\nCase 4. ",
"Maximum automation for localization\n\nDescription:\n\nCompany Y uses GitHub for file storage, its translation resources are stored in a separate repository. ",
"Crowdin automates the localization process by uploading modified strings to the project for translation. ",
"Once the translation is done and confirmed by the localization manager, the text is uploaded to the same repository with a language tag. ",
"After that, it’s automatically incorporated into the build that’s sent off for testing. ",
"The process leverages the API for multiple solutions, the company’s own platform, and a single manager.",
"\n\nResult:\n\nThat workflow delivers the translation and testing of updates at record speed regardless of their size, and it’s suited to work both with freelancers and organizations. ",
"It saves the company money, too.",
"\n\nOn the other hand, maintaining operability means regularly updating the software on your own and testing out any new functionality to make sure it fits into the process. ",
"Managing and upgrading the flow is a job for a professional localization manager, someone who knows all the nuances of every piece of software involved. ",
"And that weakness in the chain shows up when they quit, take maternity leave, or have to move on for any other reason. ",
"Finding someone to take their place can be nearly impossible (see “Assembling the team”).",
"\n\nAutomating processes within the localization department\n\nBelow you’ll find a simplified visualization of the localization process pulled from our Culturalization Guide.",
"\n\nUnlike the similar layout in Nikolai Bondarenko’s article, ours is much simpler. ",
"Today’s companies leverage automation to launch translations into all languages at the same time (the same goes for testing).",
"\n\nWe can break the automated localization process down into four segments:\n\nAutomating text import and export. ",
"For example, there are Unity extensions that import texts directly to Google Docs. ",
"Automating the localization process. ",
"There’s special translation software available for that, both server- and cloud-based. ",
"We use memoQ, Trados, Smartcat, and Crowdin depending on the client’s needs and the kind of job we’re being asked to do. ",
"Automating the link between text uploading/downloading and setting localization objectives. ",
"This tends to be the most challenging part, as game development is anything but standardized, and the range of solutions different companies use for the various dev stages can be expansive. ",
"Still, their API and ready-made cloud solutions can be leveraged to automate the localization process fairly quickly. ",
"Automating testing. ",
"It depends on the platform and other particulars, but localization testers generally go with one of the bug tracking systems out there. ",
"Jira, to take one example, is our go-to option.",
"\n\nTo learn more about automating the localization process, check out the recordings of talks given by localization managers from Wrike and Xsolla, at LocKit, the conference we host.",
"\n\nBut the simplest and best way is to have your localization contractor integrate you into the processes they already have in place. ",
"Serious companies will offer multiple ways to make that happen, giving you the strengths and weaknesses of each.",
"\n\nAnd of course, if you don’t have a task management system on your end, and you’re just sending game texts back and forth using a network drive, don’t expect to find a silver bullet that will solve all your problems.",
"\n\n5. ",
"Working with contractors"
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"Citrus pulp and wheat straw silage as an ingredient in lamb diets: effects on growth and carcass and meat quality.",
"\nTwenty \"Merinizzata Italiana\" lambs were introduced to two experimental diets. ",
"Ten animals (five males and five females, control group) received the traditional diet that is supplied by farmers in southern Italy, which comprised of oat hay ad libitum and commercial concentrate. ",
"The second group (the same number of lambs, silage group) received citrus pulp and wheat straw silage ad libitum and 70% of the commercial concentrate supplied to the control group. ",
"The animals were slaughtered after 80 days of feeding and carcass and some meat quality parameters were measured. ",
"No differences were observed for live weight between treatments, and carcass weights were similar for the two diets, but with obvious differences between sexes. ",
"Animals on silage produced carcasses with a better muscular conformation and with a lower fatness score (P<0.05). ",
"Subcutaneous fat colour was influenced by sex, being lighter in the female carcasses (P<0.05). ",
"Dissection of different anatomical parts showed a higher percentage of lean and a lower proportion of fat in silage samples compared to control ones. ",
"Ultimate pH was highly affected by sex being higher in the samples from male lambs (P<0.01), but was unaffected by diet treatment. ",
"Diet tended to affect meat shear force value which was lower in the silage samples, although, samples from all the animals were extremely tender. ",
"Meat from silage samples had a higher water content (P<0.05). ",
"Overall, in our experimental conditions, the use of citrus pulp silage seemed to be economically convenient for producing animals with substantially unmodified carcass and meat quality characteristics."
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[
" F I L E D\n United States Court of Appeals\n Tenth Circuit\n UNITED STATES COURT OF APPEALS\n OCT 6 1998\n FOR THE TENTH CIRCUIT\n PATRICK FISHER\n Clerk\n\n RICHARD WALTER JOHNSON,\n\n Petitioner-Appellant,\n\n v. No. ",
"98-1101\n (D.C. No. ",
"98-D-27)\n GALE A. NORTON, PETER (D. Colo.)\n WARREN BOOTH, GILBERT\n MARTINEZ, JOHN WESLEY\n ANDERSON, and ATTORNEY\n GENERAL FOR THE STATE\n OF COLORADO,\n\n Respondents-Appellees.",
"\n\n\n\n\n ORDER AND JUDGMENT *\n\n\n\n\nBefore ANDERSON , BARRETT , and TACHA , Circuit Judges.",
"\n\n\n\n After examining the briefs and appellate record, this panel has determined\n\nunanimously that oral argument would not materially assist the determination of\n\n\n\n\n*\n This order and judgment is not binding precedent, except under the\ndoctrines of law of the case, res judicata, and collateral estoppel. ",
"The court\ngenerally disfavors the citation of orders and judgments; nevertheless, an order\nand judgment may be cited under the terms and conditions of 10th Cir. ",
"R. 36.3.",
"\n\fthis appeal. ",
"See Fed. ",
"R. App. ",
"P. 34(a); 10th Cir. ",
"R. 34.1.9. ",
"The case is therefore\n\nordered submitted without oral argument.",
"\n\n Petitioner Richard Walter Johnson applies for a certificate of appealability\n\nso that he might appeal the district court’s dismissal of his petition for writ of\n\nhabeas corpus brought pursuant to 28 U.S.C. § 2254. ",
"He further requests\n\npermission to proceed on appeal without prepayment of costs or fees. ",
"For the\n\nreasons stated below, we deny the application for a certificate of appealability\n\nand dismiss the appeal. ",
"We further deny permission to proceed on appeal without\n\nprepayment of costs or fees.",
"\n\n Mr. Johnson is incarcerated in the El Paso County Jail in Colorado Springs,\n\nColorado. ",
"It appears from his § 2254 petition that his conviction was affirmed by\n\nthe Colorado Court of Appeals but that Mr. Johnson did not further appeal that\n\njudgment to the Colorado Supreme Court. ",
"He did file for state post-conviction\n\nrelief, but again failed to appeal the denial of that petition to the highest state\n\ncourt having jurisdiction over it. ",
"For these reasons, the district court denied\n\nMr. Johnson’s petition for failure to exhaust state remedies. ",
" See Rose v. Lundy ,\n\n455 U.S. 509, 515 (1982); see also 28 U.S.C. § 2254(b)(1).",
"\n\n We review the legal basis for the district court’s dismissal of Mr. Johnson’s\n\n§ 2254 petition de novo. ",
" See Jackson v. Shanks , 143 F.3d 1313, 1317 (10th Cir.",
"\n\n1998). ",
"After review of the materials submitted by Mr. Johnson and the record in\n\n\n -2-\n\fthis case, we find no fault with the order of the district court. ",
"Mr. Johnson’s\n\napplication for a certificate of appealability is DENIED because he has failed to\n\nmake a substantial showing of the denial of a constitutional right. ",
" See 28 U.S.C.\n\n§ 2253(c)(2). ",
"His requests to proceed on appeal without prepayment of costs or\n\nfees and for rehearing and release pending appeal are similarly DENIED.",
"\n\n This appeal is DISMISSED. ",
"The mandate shall issue forthwith.",
"\n\n\n\n Entered for the Court\n\n\n\n James E. Barrett\n Senior Circuit Judge\n\n\n\n\n -3-\n\f"
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[
"---\nabstract: 'Recently the background independent nonperturbative quantization has been extended to various theories of gravity and the corresponding quantum effective cosmology has been derived, which provides us with necessary avenue to explore the pre-inflationary dynamics. ",
"Brans-Dicke (BD) loop quantum cosmology (LQC) is one of such theories whose effective background dynamics is considered in this article. ",
"Starting with a quantum bounce, we explore the pre-inflationary dynamics of a universe sourced by a scalar field with the Starobinsky potential in BD-LQC. ",
"Our study is based on the idea that though Einstein’s and Jordan’s frames are classically equivalent up to a conformal transformation in BD theory, this is no longer true after quantization. ",
"Taking the Jordan frame as the physical one we explore in detail the bouncing scenario which is followed by a phase of a slow roll inflation. ",
"The three phases of the evolution of the universe, namely, [*bouncing, transition from quantum bounce to classical universe, and the slow roll inflation*]{}, are noted for an initially kinetic energy dominated bounce. ",
"In addition, to be consistent with observations, we also identify the allowed phase space of initial conditions that would produce at least 60 e-folds of expansion during the slow roll inflation.'",
"\nauthor:\n- 'Manabendra Sharma$^1$ [^1]'\n- 'Tao Zhu$^1$ [^2]'\n- 'Anzhong Wang$^{1,2}$ [^3]'\ntitle: 'Background dynamics of pre-inflationary scenario in Brans-Dicke loop quantum cosmology'\n---\n\nIntroduction {#Introduction}\n============\n\nThe inflationary cosmology as proposed by Alan Guth in 1981 [@Guth] provides a solution to some of the puzzles of the Standard Big Bang Cosmology by introducing an epoch of nearly exponential expansion in the early universe. ",
"Other than serving as a mechanism to solve problems like horizon, flatness and entropy etc., ",
"it is much more powerful in predicting the cosmos we see today [@Realized]. ",
"It offers a first ever causal explanation for the origin of inhomogeneities in the universe. ",
"To be precise, it predicts the primordial power spectra whose evolutions explain both the formation of the large scale structure and small inhomogeneities present in the cosmic microwave background (CMB) [@Guth; @CMB]. ",
"Today, the unprecedented success of the inflationary paradigm is based on observational precisions [@Success1]-[@Success3]. ",
"Though very successful, the paradigm still suffers from issues like initial singularity and trans-Planckian problem. ",
"For a review on conceptual issues of inflationary cosmology see [@UC].",
"\n\nFor every expanding Friedmann-Lametre-Robertson-Walker (FLRW) universe, there always appears a big-bang singularity in general relativity (GR), if matter satisfies certain energy conditions [@HE73]. ",
"This strong curvature singularity at which physics comes to halt is an artifact, because we are pushing the classical theory of gravity to a region where it is no longer valid. ",
"Mathematically speaking, the geodesics are past incomplete and hence the affine parameter cannot be extended to infinite past leading to a focusing of congruence of geodesics [@Raychaudhuri; @LargeScaleStructure]. ",
"This causes the breakdown of the notion of space-time itself. ",
"The result of focusing theorem is valid both for null and time like geodesic [@EricPoisson]. ",
"It can be shown that this issue is independent of the symmetry of the metric considered unless and until certain energy conditions are hold good and depends solely on the average value of the Hubble parameter [@MyThesis1]. ",
"Neither Standard model of cosmology nor inflationary paradigm have a say on this issue. ",
"An alternative scenario, called Bounce, within the framework of classical theory of gravity requires either the consideration of exotic stress-energy tensor or the modification of the gravity sector of the Einstein-Hilbert action which is evident from the Raychaudhuri equation in the absence of the centrifugal term [@Raychaudhuri]. ",
"In addition to solving the puzzles of the standard model of cosmology [@SolutionHPFP], these classical bouncing scenarios involve circumvention of the singularity by assuming the universe to start from a contracting phase and then bouncing back to an expanding phase well before reaching the Planck length [@MyThesis1]. ",
"However these scenarios are also faced with serious setbacks like instability problem [@Instabilities]. ",
"For a review on classical bouncing scenarios and their drawbacks see [@ReviewOnBouncing1; @ReviewOnBouncing2]. ",
"Amongst all the drawbacks the fact that the universe becomes extremely dense early on suggests that quantum gravitational effects would become important at such small length and one must adopt quantum gravity to describe the “zone of ignorance.\" ",
"Therefore, it is natural to speculate that in a true theory of quantum gravity quantum mechanics would intervene to avert the singularity.",
"\n\nLoop quantum gravity (LQG) is a candidate of quantum gravitational theory where it takes the premise of gravity as a manifestation of geometry of space-time seriously and then systematically constructs a specific theory of quantum Riemanian geometry. ",
"For a review we refer the reader to [@Thiemann1]. ",
"LQG stands out as a leading non-perturbative and background-independent approach to quantize gravity [@QuantumGravity1]. ",
"At its depth, this theory brings out a fundamental discreteness at the Planck scale wherein the underlying geometric observables, such as areas of physical surfaces and volumes of physical regions, are discrete in nature [@Asthekar1; @Asthekar2; @Bianchi1]. ",
"The avenue of loop quantum cosmology (LQC) is an application of LQG techniques to the symmetry reduced space-time, homogeneous space-time for cosmology in particular [@Bojowald1]. ",
"In LQC the singularity is resolved in the sense that physical macroscopic observables, such as energy density and curvature which diverge at the big bang in GR, have a finite upper bound. ",
"This finiteness of the macroscopic parameters above owes to the fact that they have a dependence on the microscopic parameter of the theory called the fundamental area gap whose smallest eigenvalue is nonzero [@Asthekar3; @Asthekar4; @Asthekar5]. ",
"Thus, a contracting FLRW universe would bounce back to an expanding one avoiding the formulation of singularity in LQC as there is a maximal value of energy density. ",
"This is achieved without adding any nontrivial piece of matter unlike in the case of classical bounce. ",
"This quantum bounce occurs purely due to quantum geometric effects which act as a novel repulsive force that can be easily seen from the quantum corrected Friedmann and Raychaudhuri equations to be discussed in Sec.\\[Cosmology\\]. ",
"Also, it is to be noted that in all the different classes of space-times permitting different sets of symmetries [@WE], including Bianchi and Gowdy models, the singularity is resolved in the framework of LQC [@Asthekar6]-[@Brizuela2]. ",
"Based on the loop quantization of Brans-Dick (BD) theory the dynamics of BD-LQC has also been explored [@Jin]. ",
"For a review of singularity resolution in LQC, see [@Asthekar9; @Singh3].",
"\n\nIn addition to the above, the fact that the universe must have expanded at least 50 e-folds so as to be consistent with the current observations leads to a problematic situation if the universe had expanded a little more than 70 e-folds. ",
"In fact, had it been so (which is true for a large class of inflationary models [@5]), it turns out that the wavelengths of all fluctuation modes which are currently inside the Hubble radius were smaller than the Planck length at the onset of the inflation. ",
"This is coined as the trans-Planckian issue in [@6], which challenges the validity of the assumption that matter fields are quantum in nature but spacetimes can still be treated classically which are used at the beginning of inflation in order to make predictions [@CMB]. ",
"Thus, once again it calls for quantum treatment of spacetimes. ",
"Moreover, the inflationary paradigm usually sets the adiabatic vacuum state at the time when the wavelengths of fluctuations were well within the Hubble horizon during the phase of inflation. ",
"This treatment, however, allows the ignorance of the dynamics prior to the onset of the inflation, even when the modes were well inside the Hubble horizon. ",
"For more details regarding the sensitivity of the inflationary dynamics to quantum gravitational effects we refer readers to [@6; @8; @Zhu:2013fha; @Zhu:2015xsa; @UnivFeatB].",
"\n\nAs cited above all these issues motivate to look for quantum gravity candidates and LQC stands out as, with robustness, a competing one that replaces the singularity by a quantum bounce which is followed by a desired slow roll inflation [@18]. ",
"Now LQC is in a position to undergo experimental tests and to look for observational signatures of quantum bounce, pre-inflationary dynamics in current/forth-coming observations. ",
"In fact three major streams of calculations of cosmological perturbations, namely, *dressed metric* [@Asthekar9], *deformed algebra* [@14], and *hybrid approaches* [@Mendez] have been carried out and studied numerically and analytically in [@10]-[@navascures_hybrid_2016]. ",
"It has been reported that the *deformed algebra* approach is already inconsistent with current observations [@21; @arxiv].",
"\n\nComing back to the inflationary paradigm, this standard inflationary scenario is based on a canonical scalar field in the framework of GR. ",
"Viability of different models of inflation in the framework of standard inflationary scenario have been extensively studied [@13JCAP]-[@17JCAP]. ",
"An important category of inflationary models are based on the extended theories of gravity. ",
"This classical inflationary models on the extended theories of gravity have been extensively studied in literature [@18JCAP]-[@46JCAP]. ",
"In particular the study of inflation in BD gravity has been carried out in detail in [@JCAP]. ",
"Wherein the inflationary observables containing the scalar spectral index, the tensor to scalar ratio, the running of the spectral index and the equilateral non-Gaussianity parameters in terms of general form of potential in the Jordan frame are obtained and results are compared in the light of Planck 2015 data [@JCAP]. ",
"However as the early universe is extremely hot, it is also important to consider the quantum gravitational effects of spacetime in the early universe. ",
"Now as mentioned above for the last two and half decades LQG has caught attention of the community and been widely investigated [@QuantumGravity1; @43ZhuTao], as it gives a background independent way to quantize GR. ",
"In this framework of LQG, it is remarkable that GR can be non-perturbatively quantized. ",
"Recently this promising loop quantization technique has been extended to theories of modified gravity, for instances, BD theory [@35ZhuTao; @36ZhuTao], metric $f(R)$ theories [@37ZhuTao; @38ZhuTao] and scalar-tensor theories [@17ofZhaAndMa]. ",
"Thus, it opens up an interesting avenue to explore the early universe when geometry is quantized beyond GR. ",
"That BD theory in Jordan’s frame is equivalent to Einstein’s frame classically up to a conformal transformation is no longer true after quantization in the two frames [@17ofZhaAndMa]. ",
"In [@Jin] the pre-inflationary dynamics of BD theory is extensively studied by taking Einstein’s frame as the physical one.",
"\n\nIn this paper we focus on studying the effective BD-LQC by taking Jordan’s frame as the physical one for a universe sourced by a scalar field with the Starobinsky potential. ",
"This paper is divided broadly into three main sections: Sec.\\[Cosmology\\], Sec.\\[CosmologyResults\\] and Sec.\\[DR\\]. ",
"In Sec.\\[Cosmology\\] we present briefly the background cosmology in BD gravity in classical set-up. ",
"Wherein we present the equations of motion of background dynamics both in Einstein’s and Jordan’s frames and emphasize that both are classically equivalent to each other up to a conformal transformation, provided that the same coordinate system is used. ",
"Sec.\\[CosmologyResults\\] contains our studies and main results of a loop quantum corrected BD cosmology sourced by a scalar field with the Starobinsky potential. ",
"This is further divided into two subsections \\[EV\\] and \\[NA\\]. ",
"In subsection \\[EV\\] we set up the stage by presenting the loop quantum corrected equations of motion for the background dynamics of BD theory in Jordan’s frame. ",
"We analyze the quantum corrected Friedmann and Raychaudhuri equations in detail to see whether a quantum bounce exists or not. ",
"In addition to stating the necessary conditions, we put forward an effective definition of pressure. ",
"In the subsection \\[NA\\] we present all our numerical results for a kinetic dominated bouncing scenario. ",
"Wherein we check whether a slow roll inflation is attained and subsequently calculate the number of e-folds of expansion generated for different values of the field at the quantum bounce. ",
"Finally we discuss and conclude our work in \\[DR\\].",
"\n\nDynamics of the Background Cosmology {#Cosmology}\n====================================\n\nClassical dynamics of slow roll inflation in BD theory\n------------------------------------------------------\n\nBD proposed a specific form of the scalar tensor gravity, that is based on the Mach principle, which implies that the inertial mass of an object depends on the matter distribution in the universe [@59JCAP]. ",
"The governing action for four dimensional BD theory is given by [@59JCAP], $$S_J=\\int d^4x \\sqrt{-g} \\left[ \\frac{M_{Pl}}{2} \\phi R + \\frac{M_{Pl}}{\\phi}w_{BD}X-V(\\phi)\\right],$$ where $g$ is the determinant of the spacetime metric $g_{\\mu \\nu}$, $R$ is the four dimensional Ricci scalar, $\\phi$ the BD scalar field, $w_{BD}$ the dimensionless BD parameter, whereas $X$ is defined as $X\\equiv -\\frac{1}{2} g^{\\mu \\nu} (\\partial_\\mu \\phi ) (\\partial_\\nu \\phi)$, and $V(\\phi)$ represents the potential of the scalar field $\\phi$. In this work we adopt the FLRW metric and in contrary to the original BD theory we introduce the field potential $V(\\phi)$. It is to be noted that the reduced Planck mass $M_{Pl}$ is defined as $M_{Pl}^{-2}=8 \\pi G$ and the Planck mass $m_{Pl}$ is defined as $m_{Pl}^{-2}=G$.\n\nNow $f(R)$ gravity with the action $$S= \\int d^4 x \\sqrt{-g} \\frac{M_{Pl}^2}{2} f(R),$$ can be easily obtained from BD theory if we consider the following correspondence, $$\\frac{\\phi}{M_{Pl}}=\\frac{{\\rm d} f}{{\\rm d} R},~~V(\\phi)= \\frac{M_{Pl}^2}{2}\\left( R \\frac{{\\rm d} f}{{\\rm d}R}-f \\right),~~w_{BD}=0.$$ This in turn gives the Starobinsky inflation with the action [@CMB] $$S_{R^2}= \\int d^4x \\sqrt{-g} \\frac{M_{Pl}^2}{2}\\left( R + \\frac{R^2}{6M^2} \\right), \\label{Action}$$ as a particular case of BD theory with the following transformation, $$\\frac{\\phi}{M_{Pl}}= 1+ \\frac{R}{3M^2},~~V(\\phi)=\\frac{3M^2}{4}(\\phi-M_{Pl})^2,~~w_{BD}=0.$$ Now to get the equation of motion of the background, we invoke a spatially flat, homogeneous and isotropic metric in the Jordan frame which takes the form $$ds^2=-dt^2 + a^2(t) \\delta_{ij} dx^{i}dx^{j},$$ where $a(t)$ is the scale factor of the universe and $t$ is the cosmic time defined in Jordan frame. ",
"The background dynamics of the universe, can be obtained by varying the action w.r.t the metric $g_{\\mu \\nu}$ and $\\phi$ [@49JCAP; @69JCAP] to give the modified Friedmann and Klein-Gordon equations, $$\\begin{aligned}\n3 M_{Pl}^2\\left( H + \\frac{\\dot{\\phi}}{2 \\phi} \\right)^2= \\frac{M_{Pl}^2 \\rho_{\\phi}}{\\phi^2},\\\\\n\\ddot{\\phi} + 3H\\dot{\\phi} + \\frac{2}{\\beta M_{Pl}}\\left[\\phi V_{\\phi}(\\phi) -2 V(\\phi)\\right]=0,\\end{aligned}$$ where $H\\equiv {\\dot{a}}/{a}$ is the Hubble parameter, $\\beta= 2 w_{BD} +3 $ and $\\rho_{\\phi}$ is defined as $\\rho_{\\phi}\\equiv ({\\beta}/{4}) \\dot{\\phi}^2 + {\\phi V(\\phi)}/{M_{Pl}}$ is the effective energy density of the BD scalar field. ",
"It should be noticed that for the Starobinsky inflation we have $\\beta=3.$\n\nStarobinsky potential in Einstein frame\n---------------------------------------\n\nIn this subsection we present, for the sake of completeness, the classical equation of motion of the background universe sourced by the Starobinsky potential in the Einstein’s frame. ",
"For detailed analysis of Starobinsky inflation in Einstein’s frame with loop quantum correcton see [@Jin]. ",
"Classically, the Jordan and the Einstein frames are equivalent to each other up to a conformal transformation when the same coordinate system is employed. ",
"Thus under a conformal transformation $$\\hat{g}_{\\mu \\nu} = \\frac{\\phi}{M_{Pl}}g_{\\mu \\nu}, \\label{CT}$$ the action Eq.(\\[Action\\]) can be modeled to that of a minimally coupled scalar field $\\chi$ in the Einstein frame where we use a hat to represent quantities in this frame. ",
"Thus, under this conformal transformation the action gets recasted into $$S_E= \\int d^4x \\sqrt{-\\hat{g}} \\left[ \\frac{M_{Pl}^2}{2}\\hat{R}-\\frac{1}{2}\\hat{g}_{\\mu \\nu} \\partial_{\\mu}\\chi \\partial_\\nu \\chi - U(\\chi) \\right],$$ and we can easily recognise that $$\\begin{aligned}\nU(\\chi)&=& e^{-2 \\sqrt{\\frac{2}{\\beta}}\\frac{\\chi}{M_{Pl}}}V(\\phi),\\\\\n\\frac{\\phi}{M_{Pl}} &=& e^{\\sqrt{\\frac{2}{\\beta}}\\frac{\\chi}{M_{Pl}}}.\\end{aligned}$$ Now, for the Starobinsky inflation the correspondence between scalar field $\\chi$ and the Ricci scalar is given by $$\\frac{\\chi}{M_{Pl}}= \\sqrt{\\frac{3}{2}}ln\\left( \\frac{\\phi}{M_{Pl}} \\right)=\\sqrt{\\frac{3}{2}}ln\\left( 1+ \\frac{R}{3M^2} \\right),$$ which gives $$\\begin{aligned}\n\\nonumber\nU(\\chi) = \\frac{3}{4} M^2 M_{Pl}^2 \\left( 1- e^{-\\sqrt{\\frac{2}{\\beta}}\\frac{\\chi^2}{M_{Pl}^2}} \\right),\\;\\;\\;\n\\beta = 3. ",
"\\end{aligned}$$ Note that the Starobinsky potential $U(\\chi)$ with $\\beta=3$ can be extended to a subclass of E type $\\alpha$ attractor inflationary models [@8ZhuTao; @9ZhuTao; @19JCAP]. ",
"Now considering a universe with the flat FLRW metric in the Einstein frame $$ds^2= -d\\hat{t}^2 + \\hat{a}^2 \\delta_{ij}d{x}^i d{x}^j,$$ where $\\hat{a}(\\hat{t})$, $d{x}^i$ being the scale factor and comoving coordinates in Einstein’s frame. ",
"The background dynamics, in this frame, are given by the modified Friedmann and Klein-Gordon equations $$\\begin{aligned}\n\\hat{H}^2 = \\frac{1}{3 M_{Pl}^2} \\hat{\\rho}_{\\chi}, \\\\\n\\frac{{\\rm d}^2 \\chi}{{\\rm d}\\hat t^2}+ 3 \\hat{H} \\frac{{\\rm d} \\chi}{ {\\rm d} \\hat t} + U(\\chi) &=& 0,\\end{aligned}$$ where the effective energy density is given by $\\hat{\\rho}_{\\chi}= \\frac{1}{2} \\dot{\\chi}^2 + U(\\chi)$.\n\nEquivalence between the Einstein and the Jordan frame\n-----------------------------------------------------\n\nAs has been stated above the Einstein and Jordan frames are equivalent upto a conformal transformation at the classical level. ",
"Under this transformation (\\[CT\\]) the relationship between the corresponding dynamical quantities of the theory in the two frames can be immidiately established to be $$\\hat{a}(\\hat{t}) = \\sqrt{\\frac{\\phi}{M_{Pl}}}a(t), ~~ {\\rm d} \\hat{t} = \\sqrt{\\frac{\\phi}{M_{Pl}}} {\\rm d} t,$$ for the scale factors and the cosmic times. ",
"With these the following relations can be easily found $$\\begin{aligned}\n\\hat{H} &=& \\sqrt{\\frac{M_{Pl}}{\\phi}} \\left( H + \\frac{\\dot{\\phi}}{2 \\phi} \\right),\\\\\n\\frac{{\\rm d}\\chi}{{\\rm d}\\hat{t}} &=& \\sqrt{\\frac{\\beta}{2}} \\left( \\frac{M_{Pl}}{\\phi} \\right)^{\\frac{3}{2}} \\dot{\\phi},\\\\\n\\frac{{\\rm d}^2 \\chi}{{\\rm d} \\hat{t}^2} &=& \\sqrt{\\frac{\\beta}{2}}M_{Pl}^2 \\left( \\frac{\\ddot{\\phi}}{\\phi}^2 -\\frac{3}{2} \\frac{\\dot{\\phi}^2}{\\phi^3} \\right),\\\\\n\\hat{\\rho}_{\\chi} &=& \\left( \\frac{M_{Pl}}{\\phi} \\right)^3 \\left[ \\frac{\\beta}{4} \\dot{\\phi}^2 + \\frac{\\phi}{M_{Pl}} V(\\phi) \\right].\\end{aligned}$$ Note that though these transformation between the two frames hold good at the classical level, it does not turn out to be so after quantization. ",
"In fact the effective dynamics is different and this point is what we explore in this work.",
"\n\nEffective background dynamics of Brans-Dicke loop quantum cosmology {#CosmologyResults}\n===================================================================\n\nEffective background dynamics of BD-LQC in Jordan’s frame {#EV}\n---------------------------------------------------------\n\nIn this subsection we present the required equations to numerically simulate the effective quantum corrected background dynamics in BD-LQC in Jordan’s frame. ",
"The modified Friedmann equation and the Klein-Gordon equation are sufficient to study the background dynamics. ",
"The quantum corrected Friedmann equation for the effective BD-LQC theory in Jordan frame is [@36ZhuTao] $$\\left( H + \\frac{\\dot{\\phi}}{2\\phi}\\right)^2 = \\left[ \\frac{1}{\\phi}\\sqrt{\\frac{\\rho_{\\phi}}{3}}\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}} + \\frac{\\dot{\\phi}}{2\\phi} \\left( 1-\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}}\\right) \\right]^2, \\label{FD}$$ where the effective energy density is given by $\\rho_{\\phi}= \\frac{\\beta}{4} \\dot{\\phi}^2 + \\frac{\\phi V(\\phi)}{M_{Pl}}$. The quantum corrected Klein-Gordon equation for BD theory in Jordan’s frame is $$\\ddot{\\phi} + 3H \\dot{\\phi} + \\frac{2}{\\beta M_{Pl}} \\phi V_{\\phi} + \\frac{2}{\\beta M_{Pl}} V(\\phi)\\left( 1- 3 \\sqrt{1- \\frac{\\rho_{\\phi}}{\\rho_c}} \\right)=0. ",
"\\label{KG}$$ As stated above, we need to solve these two equations to explore the background dynamics. ",
"Eq.(\\[FD\\]) immediately gives $$\\left( H + \\frac{\\dot{\\phi}}{2\\phi}\\right) = \\pm \\left[ \\frac{1}{\\phi}\\sqrt{\\frac{\\rho_{\\phi}}{3}}\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}} + \\frac{\\dot{\\phi}}{2\\phi} \\left( 1-\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}}\\right) \\right]. ",
"\\label{FD1}$$ Thus we get two solutions. ",
"The $``+\"$ solution of Eq.(\\[FD1\\]) gives $$H= \\frac{1}{\\phi}\\left[ \\sqrt{\\frac{\\rho_{\\phi}}{3}} -\\frac{\\dot{\\phi}}{2}\\right]\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}}, \\label{H1}$$ and the $``-\"$ solution gives $$H = -\\frac{\\dot{\\phi}}{\\phi} - \\frac{1}{\\phi}\\left( \\sqrt{\\frac{\\rho_{\\phi}}{3}} -\\frac{\\dot{\\phi}}{2}\\right) \\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}}. ",
"\\label{H2}$$ Before defining some useful parameters to carry out qualitative and quantitative analysis of our models considered in this work, let us check if we attain a nonsingular bouncing model in each of theses two cases, namely, given by, Eq.(\\[H1\\]) and Eq.(\\[H2\\]), respectively.",
"\n\nA nonsingular bounce is obtained as when an initially contracting universe goes to an expanding one through a minimum in the scale factor but not zero. ",
"Since the Hubble parameter is proportional to the fractional rate of change of volume, $H \\propto \\frac{\\triangle V}{V \\triangle t}$, therefore a contracting (expanding) phase is specified by a negative (positive) value of the Hubble parameter H. However this is a necessary but not sufficient condition for bounce to occur. ",
"Along with this the slope of the slope of the scale factor must be positive at the turn around to achieve the minimum and hence the bounce. ",
"Mathematically speaking the following two conditions must be met for the bounce to occur, $$\\begin{aligned}\n\\left(\\frac{{\\rm d}a}{{\\rm d}t}\\right)_B=0,\\label{Condition1} \\\\ \n\\left( \\frac{{\\rm d}^2 a}{{\\rm d}t^2} \\right)_B > 0. ",
"\\label{Condition2}\\end{aligned}$$ From here onwards the subscript B will be used to denote the point of the bounce. ",
"Now let us examine if the two conditions given by Eqs.(\\[Condition1\\]) and (\\[Condition2\\]) are satisfied for the two solutions of Eq.(\\[H1\\]) (for the $``+\"$ solution) and Eq.(\\[H2\\]) (for the $``-\"$ solution) respectively. ",
"In the contracting phase, as the volume of the universe decreases and therefore the energy density of the universe increases, we expect the universe to turn around when it reaches the maximal energy scale $\\rho_c$ given by the theory due to quantum geometric effects. ",
"Thus, we need first to check if the Hubble parameter is zero when $\\rho_{\\phi}= \\rho_{c}$. From Eq.(\\[H1\\]) it is clear that $$H=\\frac{1}{\\phi}\\left( \\sqrt{\\frac{\\rho_{\\phi}}{3}} -\\frac{\\dot{\\phi}}{2}\\right)\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_{c}}}= 0 ~~~at~~ \\rho_{\\phi}=\\rho_{c},$$ and for Eq.(\\[H2\\]) we find $$H=-\\frac{\\dot{\\phi}}{\\phi} -\\frac{1}{\\phi}\\left( \\sqrt{\\frac{\\rho_{\\phi}}{3}} -\\frac{\\dot{\\phi}}{2}\\right)\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_{c}}}= 0 ~~~at~~ \\rho_{\\phi}=\\rho_{c} ~~if ~and~ only~ if ~~\\dot{\\phi}|_{\\rho_{\\phi}=\\rho_c}.$$ But with these, only Eq.(\\[Condition1\\]) is fulfilled. ",
"We need to see explicitly for each of the solution of H whether the second condition Eq.(\\[Condition2\\]) is also satisfied or not (at $\\rho_{\\phi}=\\rho_{c})$. As we know that given the Friedmann and Klein-Gordon equations, the Raychaudhuri equation can be derived using the following identity $$\\frac{\\ddot{a}}{a} = H^2 + \\frac{{\\rm d}H}{{\\rm d}t}.$$ Now let us derive it for each of the equations (\\[H1\\]) and (\\[H2\\]) case by case. ",
"Considering first Eq.(\\[H1\\]), as we have seen above that the Hubble parameter becomes zero as $\\rho_{\\phi}=\\rho_c$, therefore, $$\\left( \\frac{\\dot{a}}{a} \\right)_{\\rho_{\\phi}=\\rho_{c}} = \\left( \\frac{{\\rm d} H}{{\\rm d} t} \\right)_{\\rho_{\\phi}=\\rho_{c}}.$$ Then we find $$\\begin{aligned}\n\\frac{{\\rm d}H}{{\\rm d}t}&=& \\frac{{\\rm d}}{{\\rm d}t} \\left[ \\frac{1}{\\phi}\\left[ \\frac{\\rho_{\\phi}}{3} -\\frac{\\dot{\\phi}}{2}\\right]\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}} \\right],\\\\\n &=& \\frac{{\\rm d}}{{\\rm d}t} \\left[ \\frac{1}{\\phi}\\left( \\frac{\\rho_{\\phi}}{3} -\\frac{\\dot{\\phi}}{2}\\right)\\right]\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}} ~+~ \\frac{1}{2\\phi}\\left(\\sqrt{\\frac{\\rho_{\\phi}}{3}}-\\frac{\\dot{\\phi}}{2} \\right) \\frac{-\\frac{1}{\\rho_c}\\frac{{\\rm d}\\rho_{\\phi}}{{\\rm d}t} }{\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_{c}}}}.\\end{aligned}$$ Evaluating the above expression at $\\rho_{\\phi}= \\rho_{c}$, we are left with only the second term $$\\left(\\frac{{\\rm d}H}{{\\rm d}t}\\right)_{\\rho_{\\phi}=\\rho_c}=\\left( \\frac{1}{2\\phi}\\left(\\sqrt{\\frac{\\rho_{\\phi}}{3}}-\\frac{\\dot{\\phi}}{2} \\right) \\frac{-\\frac{1}{\\rho_c}\\frac{{\\rm d}\\rho_{\\phi}}{{\\rm d}t} }{\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_{c}}}} \\right)_{\\rho_{\\phi}=\\rho_{c}}.$$ Care must be taken while taking the limit. ",
"Note that first we need to evaluate the differentiation and then take the limit. ",
"In order to do that let us begin with the definition of $\\rho_{\\phi}= \\frac{\\beta}{4} \\dot{\\phi}^2 + \\frac{\\phi V(\\phi)}{M_{Pl}}$. Taking the differentiation w.r.t. ",
"time we have $$\\frac{{\\rm d}\\rho_{\\phi}}{{\\rm d}t}= \\left[ \\frac{\\beta}{2}\\ddot{\\phi} + \\frac{1}{M_{Pl}}\\left( V(\\phi)+ \\phi V_{\\phi}\\right)\\right]\\dot{\\phi}.$$ As obvious, the scalar field must satisfy the Klein-Gordon equation (\\[KG\\]). ",
"Thus using Eq.(\\[KG\\]) we have $$\\frac{\\beta}{2}\\ddot{\\phi}= -3H\\frac{\\beta}{2}\\dot{\\phi}-\\frac{\\phi V_{\\phi}}{M_{Pl}} -\\frac{V(\\phi)}{M_{Pl}} + \\frac{3V(\\phi)}{M_{Pl}}\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}}.$$ Substituting $\\frac{\\beta}{2}\\ddot{\\phi}$ in the expression for $\\frac{{\\rm d} \\rho_{\\phi}}{{\\rm d}t}$ given above we get, $$\\frac{{\\rm d} \\rho_{\\phi}}{{\\rm d}t}= \\left[ -3H\\frac{\\beta}{2}\\dot{\\phi} + \\frac{3V(\\phi)}{M_{Pl}} \\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}}\\right] \\dot{\\phi}.$$ As at the present we are looking for deriving the expression for the Raychaudhuri equation of the dynamics dictated by Eq.(\\[H1\\]), the Hubble parameter in the above expression must be replaced from Eq.(\\[H1\\]) to get $$\\begin{aligned}\n\\frac{{\\rm d}\\rho_{\\phi}}{{\\rm d}t}&=& \\dot{\\phi}\\left[-3\\frac{\\beta}{2}\\frac{\\dot{\\phi}}{\\phi} \\left(\\sqrt{\\frac{\\rho_{\\phi}}{3}}-\\frac{\\dot{\\phi}}{2}\\right) + 3 \\frac{V(\\phi)}{M_{Pl}} \\right]\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}},\\\\\n&=& -3\\frac{\\dot{\\phi}}{\\phi} \\left[ \\frac{\\sqrt{3}}{2}\\dot{\\phi}\\sqrt{\\rho_{\\phi}} -\\rho_{\\phi} \\right]\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}},\\end{aligned}$$ where we have used the definition of $\\rho_{\\phi}$ and the fact that $\\beta=3$ to arrive at the above expression. ",
"Now substituting $\\frac{{\\rm d}\\rho_{\\phi}}{{\\rm d}t}$ back into $\\frac{{\\rm d}H}{{\\rm d} t}$, we have $$\\left(\\frac{{\\rm d}H}{{\\rm d} t}\\right)_{\\rho_{\\phi}=\\rho_{c}}= \\frac{3}{2} \\left[ \\left( \\frac{\\dot{\\phi}}{\\phi^2} \\right)\\left( \\frac{1}{\\rho_c}\\right)\\left(\\sqrt{\\frac{\\rho_{\\phi}}{3}} - \\frac{\\dot{\\phi}}{2} \\right) \\left( \\frac{\\sqrt{3}}{2} \\dot{\\phi}\\sqrt{\\rho_{\\phi}}- \\rho_{\\phi}\\right) \\right]_{\\rho_{\\phi}=\\rho_{c}}.\\label{using}$$ Now as $\\rho_{\\phi}=\\frac{\\beta}{4}\\dot{\\phi}^2+\\frac{\\phi V(\\phi)}{M_{Pl}}$, therefore, $\\sqrt{\\frac{\\rho_{\\phi}}{3}} = \\sqrt{ \\frac{\\dot{\\phi}^2}{4} + \\frac{\\phi V(\\phi)}{3 M_{Pl}}}$ will always be greater than $\\frac{\\dot{\\phi}}{2}$ for positive valued potential with $\\beta=3$ as in our case. ",
"It can be easily seen that $\\frac{\\sqrt{3}}{2}\\dot{\\phi}\\sqrt{\\rho_{\\phi}}-\\rho_{\\phi}$ is always less than zero. ",
"Thus we arrive at the conclusion that $$\\left(\\frac{{\\rm d}^2 a}{{\\rm d}t^2}\\right)_{\\rho_{\\phi}=\\rho_c} = \\left( \\frac{{\\rm d}H}{{\\rm d}t}\\right)_{\\rho_{\\phi}=\\rho_{c}} > 0, ~~{\\mbox{only~if}} ~(\\dot{\\phi})_{\\rho_{\\phi}=\\rho_c}<0.$$ Therefore, to have $\\rho_{\\phi}=\\rho_{c}$ as the bouncing point, only the locus of negative initial velocity of the field are allowed. ",
"Thus, in this work we consider only the case with $\\dot{\\phi}_B<0$ as it is the only allowed initial value of the velocity of the scalar field $\\phi$ for bounce to happen at $\\rho_{\\phi}=\\rho_{c}$ for the dynamics given by Eq.(\\[H1\\]).",
"\n\nNext, we shall examine if $\\rho_{\\phi}=\\rho_{c}$ is also a bouncing solution for Eq.(\\[H2\\]). ",
"We begin as above with the identity $\\frac{\\ddot{a}}{a}= \\frac{{\\rm d}H}{{\\rm d}t} +H^2$ but now with $H$ given by Eq.(\\[H2\\]). ",
"From Eq.(\\[H2\\]) we have seen that the first condition Eq.(\\[Condition1\\]) is satisfied for $\\rho_{\\phi}=\\rho_{c}$ provided that $(\\dot{\\phi})_{\\rho_{\\phi}=\\rho_c}=0$. This is a very special case. ",
"Now let us examine if this initial condition $(\\dot{\\phi})_{\\rho_{\\phi}=\\rho_{c}}=0$ also satisfies the second condition for a bounce to occur. ",
"This amounts to check if $$\\left( \\frac{\\ddot{a}}{a} \\right)_{\\rho_{\\phi}=\\rho_{c}} = \\left(\\frac{{\\rm d}H}{{\\rm d}t}\\right)_{\\rho_{\\phi}=\\rho_{c}},$$ is greater than zero for $(\\dot{\\phi})_{\\rho_{\\phi}=\\rho_{c}}=0$. Now $\\frac{{\\rm d}H}{{\\rm d}t}$ for the dynamics given by Eq.(\\[H2\\]) is $$\\frac{{\\rm d}H}{{\\rm d}t}=-\\frac{{\\rm d}}{{\\rm d}t}\\left(\\frac{\\dot{\\phi}}{\\phi}\\right) - \\frac{{\\rm d}}{{\\rm d}t}\\left[\\frac{1}{\\phi} \\left(\\sqrt{\\frac{\\rho_{\\phi}}{3}} -\\frac{\\dot{\\phi}}{2} \\right)\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}} \\right].$$ The second term on the right hand side of the above equation is already evaluated above while analyzing the fulfillment of second bouncing condition for Eq.(\\[H1\\]) (for $``+\"$ solution of H) and hence using Eq.(\\[using\\]) the above equation becomes $$\\frac{{\\rm d}H}{{\\rm d}t}= -\\frac{\\ddot{\\phi}}{\\phi} + \\left(\\frac{\\dot{\\phi}}{\\phi}\\right)^2 + \\frac{3}{2} \\left( \\frac{\\dot{\\phi}}{\\phi} \\right)\\left( \\frac{1}{\\rho_c} \\right) \\left( \\sqrt{\\frac{\\rho_{\\phi}}{3}}- \\frac{\\dot{\\phi}}{2}\\right) \\left( - \\frac{\\sqrt{3}}{2}\\sqrt{\\rho_{\\phi}} + \\rho_{\\phi} \\right).$$ But at $\\rho_{\\phi}=\\rho_{c}$ the initial value of the velocity of the scalar field must be zero to satisfy the first condition of bounce. ",
"Therefore, we have $$\\left(\\frac{{\\rm d}H}{{\\rm d}t}\\right)_{\\rho_{\\phi}=\\rho_{c}}= -\\left(\\frac{\\ddot{\\phi}}{\\phi}\\right)_{\\rho_{\\phi}=\\rho_c}.$$ Now to check whether the second condition of a bounce is satisfied or not, all we have to do is to evaluate the quantity $-\\left(\\frac{\\ddot{\\phi}}{\\phi} \\right)_{\\rho_{\\phi}=\\rho_c}$ from the Klein-Gordon equation (\\[KG\\]) to have, $$-\\frac{\\ddot{\\phi}}{\\phi}= + \\frac{3H\\dot{\\phi}}{\\phi} + \\frac{2}{\\beta M_{Pl}}\\frac{\\phi V_{\\phi}}{\\phi} + \\frac{2}{\\beta M_{Pl}}\\frac{V(\\phi)}{\\phi} -\\frac{2}{\\beta M_{Pl}} \\frac{V(\\phi)}{\\phi}3\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_{c}}},$$ and evaluating the above at $\\rho_{\\phi}=\\rho_c$ we have $$-\\left( \\frac{\\ddot{\\phi}}{\\phi}\\right)_{\\rho_{\\phi}=\\rho_c}= \\frac{2}{\\beta M_{Pl}}\\left[ V_{\\phi} + \\frac{V(\\phi)}{\\phi} \\right].$$ It can be easily seen that for $\\phi$ greater than $M_{Pl}$, we have $\\left(\\frac{\\ddot{a}}{a}\\right)_{\\rho_{\\phi}=\\rho_{c}} > 0$, which gives a bouncing solution for the dynamics given by Eq.(\\[H2\\]). ",
"Also, tNote that the quantum bounce occurs only for one value of the initial condition given by $\\dot{\\phi}_B=0$. Although this is possible, it is a very special case in which the kinetic energy at the bounce to be zero. ",
"Since in this paper we are mainly concerned with the case in which the kinetic energy dominates at the bounce, we shall not consider this case further.",
"\n\nNow let us turn to study the background dynamics, for which we introduce a few parameters.",
"\n\n\\(1) The Effective equation of state (EOS), $w(\\phi)$: It is defined as the ratio of pressure to energy density $$w(\\phi)= \\frac{P_{\\phi}}{\\rho_{\\phi}}.$$ As we have an effective definition of energy density which is given by $\\rho_{\\phi}= \\frac{\\beta}{4}\\dot{\\phi}^2+ \\frac{\\phi V(\\phi)}{M_{Pl}}$, an effective definition of pressure of the scalar field $P_{\\phi}$ can always be defined if we demand that $P_{\\phi}$ which satisfy the continuity equation $$\\frac{{\\rm d} \\rho_{\\phi}}{{\\rm d}t} + 3H\\left(\\rho_{\\phi}+ P_{\\phi}\\right)=0.$$ Now as the scalar field must satisfy the Klein-Gordon equation, using Eq.(\\[KG\\]) an effective expression of pressure can be obtained to be as follows. ",
"From the definition of the effective energy density we have $$\\frac{{\\rm d} \\rho_{\\phi}}{{\\rm d}t} = \\dot{\\phi} \\left[ \\frac{\\beta}{2}\\ddot{\\phi} + \\frac{1}{M_{Pl}} \\left( V(\\phi)+ \\phi V_{\\phi}\\right) \\right],$$ and from the continuity equation we have $$\\begin{aligned}\nP_{\\phi} &=& -\\frac{1}{3H}\\frac{{\\rm d} \\rho_{\\phi}}{{\\rm d} t} - \\rho_{\\phi}\\\\ \n &=& -\\frac{1}{3H} \\dot{\\phi} \\left[ \\frac{\\beta}{2}\\ddot{\\phi} + \\frac{1}{M_{Pl}} \\left( V(\\phi)+ \\phi V_{\\phi}\\right) \\right]- \\rho_{\\phi}. ",
" \\end{aligned}$$ As the scalar field must satisfy the Klein-Gordon equation, therefore, using Eq.(\\[KG\\]) we can write $\\ddot{\\phi}$ as, $$\\begin{aligned}\n\\ddot{\\phi} &=& -3H\\dot{\\phi} - \\frac{2}{\\beta M_{Pl}}\\phi V_{\\phi}- \\frac{2}{\\beta M_{Pl}}V(\\phi) + \\frac{6}{\\beta M_{Pl}} V(\\phi) \\sqrt{1- \\frac{\\rho_{\\phi}}{\\rho_{c}}}, \\\\\n \\Rightarrow \\frac{\\beta}{2} \\ddot{\\phi} &=& -3H\\left(\\frac{\\beta}{2} \\right)\\dot{\\phi} -\\frac{\\phi V_{\\phi}}{M_{Pl}} - \\frac{V(\\phi)}{M_{Pl}} + 3\\frac{V(\\phi)}{M_{Pl}}\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_{c}}}.\\end{aligned}$$ Substituting it into the above equation for $P_{\\phi}$ we get $$P_{\\phi}= \\frac{\\beta}{4}\\dot{\\phi}^2 - \\frac{\\phi V(\\phi)}{M_{Pl}} - \\frac{V(\\phi) \\dot{\\phi}}{H M_{Pl}}\\sqrt{1-\\frac{\\rho_{\\phi}}{\\rho_c}}.$$ Now since we are considering the dynamics of the background given by Eq.(\\[H1\\]), let us substitute the Hubble parameter given by Eq.(\\[H1\\]) to get explicitly the effective pressure in terms of $\\phi$ and $\\dot{\\phi}$ $$\\begin{aligned}\n\\nonumber\nP_{\\phi} &=& \\frac{\\beta}{4} \\dot{\\phi}^2 - \\frac{\\phi V(\\phi)}{M_{Pl}} - \\frac{V(\\phi) \\dot{\\phi} \\phi}{M_{Pl} \\left(\\sqrt{\\frac{\\rho_{\\phi}}{3}} -\\frac{\\dot{\\phi}}{2} \\right)}{\\nonumber}\\\\\n&=& \\frac{\\beta}{4}\\dot{\\phi}^2 - \\frac{\\phi V(\\phi)}{M_{Pl}} - 3\\dot{\\phi} \\left[ \\sqrt{\\frac{1}{3}\\left( \\frac{\\beta}{4}\\dot{\\phi}^2 + \\frac{\\phi V(\\phi)}{M_{Pl}}\\right)}+\\frac{\\dot{\\phi}}{2} \\right].\\end{aligned}$$ Therefore, the effective equation of state parameter $w(\\phi)$ is given by $$w(\\phi) = \\frac{P_{\\phi}}{\\rho_{\\phi}} = \\frac{\\frac{\\beta}{4}\\dot{\\phi}^2 - \\frac{\\phi V(\\phi)}{M_{Pl}} - 3\\dot{\\phi} \\left[ \\sqrt{\\frac{1}{3}\\left( \\frac{\\beta}{4}\\dot{\\phi}^2 + \\frac{\\phi V(\\phi)}{M_{Pl}}\\right)}+\\frac{\\dot{\\phi}}{2} \\right]}{\\frac{\\beta}{4}\\dot{\\phi}^2 + \\frac{\\phi V(\\phi)}{M_{Pl}}}.$$\n\n\\(2) The inflation is characterized by acceleration of the universe $\\ddot{a}>0$. However it is important to have a prolonged period of inflation to produce enough number of e-folds to cure the problems of Big Bang cosmology. ",
"This is often characterized by slow roll parameters. ",
"In BD theory with the Starobinsky potential we have three slow roll parameters $$\\epsilon_1 = -\\frac{\\dot{H}}{H^2}, ~~\\epsilon_2 = \\frac{\\ddot{\\phi}}{H\\dot{\\phi}}, ~~\\epsilon_3=\\frac{\\dot{\\phi}}{2H\\phi}.$$ The slow roll inflation is achieved when $\\epsilon_1,~\\epsilon_2, \\epsilon_3\\ll 1$, and it ceases if any of them becomes 1 [@JCAP].",
"\n\n\\(3) The number of e-folds, $N_{inf}$: The amount of expansion of the universe during inflation is characterized as $N_{inf}=ln(\\frac{a_f}{a_i})=\\int^{t_f}_{t_i} Hdt\\simeq \\int^{\\phi_i}_{\\phi_f} \\frac{H}{\\dot{\\phi}} d\\phi$. During the phase of the slow roll inflation, using the slow roll condition, this can be readily seen to be $$N_{inf}= ln\\left(\\frac{a_f}{a_i} \\right) \\simeq \\int^{\\phi_i}_{\\phi_{end}} \\frac{V}{V'(\\phi)} d\\phi.$$ In this work, we will use the subscript “i\" and “f\" to denote, respectively, the starting and end times of the slow roll inflation. ",
"We use $\\ddot{a}(t=t_i)=0$ to get the starting time of inflation, whereas $\\epsilon_1$ to obtain the end time of inflation. ",
"Though strictly speaking this choice of starting time of inflation is not in accordance with the definition of the slow roll inflation, nevertheless, it is safe to use them because this is insensitive to observations [@o; @UnivFeatA]. ",
"As the fulfillment of slow roll condition only demands the slow roll parameters to be very very less than one, therefore, there is not a precise convention for the start time. ",
"In Ref.[@o; @UnivFeatA], it has been shown numerically that the change in the value of $N_{inf}$ due to different choices of start time is $0.2$ for the Starobinsky potential which is negligible for all practical purposes. ",
"To support our argument further we refer the reader to [@ToSupport], wherein authors have shown for the end time of inflation the choice of Hubble slow roll parameter or potential slow roll parameter is irrelevant for all practical purposes.",
"\n\n\\(4) An useful parameter to quantify the dominance of either kinetic energy or potential energy is via $r_{w}\\equiv \\frac{\\dot{\\phi}^2/2}{V(\\phi)}$. It is obvious that for the case of the potential energy dominant (PED) case at the bounce $r_{w}<1$, whereas it is $r_{w}>1$ for kinetic energy dominant (KED) case. ",
"In particular, we are interested in obtaining the critical value $r_w^c$ that would generate 60 e-folds of expansion of the universe in the slow roll regime.",
"\n\nNumerical Analysis {#NA}\n------------------\n\nIn this subsection we present our numerical results of the background dynamics for a quantum corrected BD cosmology in the Jordan frame for the Starobinsky potential. ",
"To study the evolution of the background universe it is sufficient to consider the quantum corrected Friedmann equation (\\[FD\\]) and the Klein-Gordon equation (\\[KG\\]) as the Raychaudhuri equation can be obtained from these and hence redundant. ",
"To begin our numerical evolution we note that the space of the initial data is four dimensional. ",
"This includes the initial value of the scale factor $a$, the Hubble parameter $H$, the scalar field $\\phi$ and its velocity $\\dot{\\phi}$. However the value of the scale factor at the initial time enjoys a constant rescaling freedom without altering the physics. ",
"Using this freedom one can fix the scale factor at bounce so that $a_B=1$ without loss of any generality. ",
"The choice of the quantum bounce as the initial condition further gives $H=0$ satisfying the first condition Eq.(\\[Condition1\\]) at $\\rho_{\\phi}=\\rho_{c}$. This in turn determines the initial value of $\\dot{\\phi}$ up to a sign once we supply the initial value of the scalar field at the bounce $\\phi_{B}$. Hence, with $a_B$ fixed the value of $\\phi_B$ and the sign of $\\dot{\\phi_B}$ completely determine the space of the initial data. ",
"In fact the set of initial conditions is the locus of all points on the phase space of $\\phi$ and $\\dot{\\phi}$ satisfying the condition $\\rho_c= \\frac{\\beta}{4}\\dot{\\phi}_B^2+ V(\\phi_B)$. In this paper we concentrate on an initially kinetic energy dominated universe with negative initial sign of $\\dot{\\phi}.$\n\nIn order to simulate the background dynamics we use Eqs.(\\[KG\\]) and (\\[H1\\]) with initial conditions as defined above. ",
"The fact that we choose Eq.(\\[H1\\]) over Eq.(\\[H2\\]) is because for dynamics dictated by Eq.(\\[H2\\]), this is a very special universe which allows only initial conditions satisfying $\\dot{\\phi_B}=0$ in order to give a bouncing solution, and this corresponds to an extremely potential dominated universe and therefore is not considered in this work.",
"\n\nThe first step to begin the numerical analysis is to calculate the mass of the Starobinsky potential in the Jordan frame, which will be simply adopted from [@JCAP], $1.4286 \\times 10^{-6}~m_{Pl}$. Thus, with the knowledge of the mass we are ready to numerically evolve the system of Eqs.(\\[KG\\]) and (\\[H1\\]) for a kinetic dominated universe initially. ",
"Let us, thus, consider three initial values of $\\phi_B/m_{Pl} = ~ (295, 395, 495)$ satisfying the initial bouncing conditions. ",
"That is, let us displace the scalar field $\\phi$ far away from the minimum of the potential and set the initial velocity of the field to be negative. ",
"Note that this negative velocity is required to fulfill the second condition of the bounce as far as dynamics given by Eq.(\\[H1\\]) is concerned. ",
"Thus, we expect that, if the field is displaced far away from the minimum and set to have a negative velocity initially, it would roll down the potential well, and it would be interesting to see when it will give rise to a slow roll inflation.",
"\n\nThe first hint of it can be seen in the plot of Fig.",
" \\[Fig1\\] where we show the variation of the scalar field $\\phi$ and its velocity $\\dot{\\phi}$ for the three initial conditions mentioned above. ",
"Note that starting with these initial conditions and with sign of the velocity being negative, eventually, the field $\\phi$ will approach the minimum of the potential and correspondingly its velocity becomes zero. ",
"Not only that the scalar field approaches the minimum of the potential for all the chosen initial values, but also the variation of the field w.r.t the cosmic time is negligible after a certain time, say $10^{4}~t/t_{Pl}$, which is a first clue that the universe might have entered a slowly roll phase of inflation. ",
"Before going to analyze whether a slow roll inflation is achieved or not, let us first look at the plot of the Hubble parameter in the lefthand side of Fig.",
" \\[Fig3\\]. ",
"It shows that starting from the bouncing point the Hubble parameter $H$ initially increases and then decreases and finally becomes almost constant. ",
"The initial rise of the Hubble parameter indicates a period of super inflation untill it reaches its maximum. ",
"Thus, similar to the models of LQC, in BD-LQC it also shows the fact that the quantum geometric effects does not merely solve the initial singularity but its novel repulsive force gives rise to a short period of super inflation $\\frac{{\\rm d} H}{{\\rm d} t}>0$.\n\nComing to the plots of the slow roll parameters $\\epsilon_1, ~ \\epsilon_2, ~ \\epsilon_3$, respectively, in Fig.",
" \\[Fig4\\] and in the left-hand side of Fig.",
" \\[Fig5\\], we can see the fulfillment of the slow roll conditions, $\\epsilon_1, ~ \\epsilon_2, ~ \\epsilon_3 \\ll 1$ in the period, $10^4 \\sim 10^7$ $t/t_{Pl}$ (for exact values of the beginning and ending of the inflation see Table \\[Table1\\]). ",
"Thus a slow roll phase of inflation is achieved. ",
"Next, our task is to calculate the number of e-folds of expansion generated during this slow roll phase of inflation for various values of the initial values of the scalar field $\\phi$ and find out what value of $\\phi_B$ gives rise to 60 e-folds of inflation. ",
"To this purpose we use $\\ddot{a}=0$ to obtain the initial time and $\\epsilon_1 = 1$ to obtain the end time of inflation. ",
"Since by definition $N_{inf}= ln(\\frac{a_f}{a_i})$, therefore, once we have the knowledge of starting and ending times of inflation, it is straintforward to carry out the calculation of $N_{inf}$ from the right plot of Fig.",
" \\[Fig2\\]. ",
"Table \\[Table1\\] shows $N_{inf}$ as a function of $\\phi_B$ for the Starobinsky potential in the Jordan frame for a kinetic energy dominated bounce with $\\dot{\\phi}_B<0$. Here, we begin with an initial value of $\\phi_B=125~m_{Pl}$ to achieve a small number of e-folds of expansion of the order of $\\simeq 4$. Since this is not adequate to solve the puzzles of the Standard Model of cosmology, which requires $\\sim 60$ e-folds of expansion, we increase in big steps the values of $\\phi_B$ and see when $60$ is attained. ",
"It is found that $60.83$ number of e-folds are generated for initial condition $\\phi_B=395m_{Pl}$ and hence it is considered as the critical value of the initial value we are seeking for and the critical energy density at the bouncing to attain this is $r_w^c = 1.15075 \\times 10^6$. Then, we continue this process to investigate the effects of $\\phi_{B}$ on $N_{inf}$ up to $\\phi_B \\simeq 1000 m_{Pl}$ and plot it in the right-hand side of Fig.",
" \\[Fig5\\]. ",
"In Fig.",
" \\[wphi\\] we plot the effective equation of state $w_{\\phi}$. Starting with a kinetic energy dominated bounce it shows the three phases of evolution, namely. [*",
"bouncing phase, transition from quantum bounce to classical universe and the slow roll inflation*]{}. ",
"During the bouncing phase as the kinetic energy dominates over potential, the effective equation of state $w_{\\phi}\\approx1$, while in the slow roll inflation phase $w_{\\phi}$ becomes $-1$, which reflects the fact that the universe is dominated by the potential energy. ",
"We also note the universality of the solution in the bouncing phase. ",
"Finally in Fig.",
" \\[Fig6\\] we plot the effective energy density due to scalar field $\\rho_{\\phi}$ on the right-hand side and the ratio $r_w$ on the left-hand side. ",
"The plot of $\\rho_{\\phi}$ in Fig.",
" \\[Fig6\\] shows the fact that the energy density becomes maximal at the quantum bounce and then dilutes away as the universe expands to large volumes as expected. ",
"Whereas the right-hand side of Fig.",
" \\[Fig6\\] demonstrates the dominance of kinetic over the potential energy throughout the evolution of the universe starting from the quantum bounce. ",
"As we initialize the universe with a kinetic energy dominated quantum bounce the ratio $r_w(t_B)\\gg1$, and then it transits to $r_w\\ll1$ as the universe becomes potential dominated in the slow roll inflation era.",
"\n\nWe have already observed that a $\\phi_B=125 m_{Pl}$ could only produce $4.49$ number of e-folds of expansion of the universe. ",
"Thus it would be interesting to see the fate of the universe for lower values of $\\phi_B$, say lower than $100 m_{Pl}$. In Fig.",
" \\[Fig7\\] we analyze this case with $\\phi_B=60,~65, ~70~m_{Pl}$ and $\\dot{\\phi_B}<0$. The evolution of $\\phi$ and $\\dot{\\phi}$ on the top panel of Fig.",
" \\[Fig7\\] suggests that the scalar field and its velocity approach to their respective minimum and start oscillating about it. ",
"This is because the inflation quickly rolls down to the minimum of the potential and undergoes damped harmonic oscillations before coming to rest in the asymptotic limit. ",
"This damped nature is because of the expansion of the universe as it can be easily seen from the Klein-Gordon equation. ",
"Thus for initial values $\\phi_B \\lesssim 100m_{Pl}$ the universe fails to undergo inflation and hence no slow roll inflation exists. ",
"As a proof of it we show the plot of $\\ddot{a}$, $\\epsilon_1$ on the bottom panel of Fig.\\[Fig7\\]. ",
"This shows that though the universe shows inflation, i.e. $\\ddot{a}>0$, it happens only for a very short period of time as it starts oscillating afterwards. ",
"Moreover, a slow roll phase of inflation is never achieved as the Hubble slow roll parameter $\\epsilon_1= -\\frac{\\dot{H}}{H^2}$ is never less than 1.",
"\n\n$\n \\begin{array}{c c}\n \\includegraphics[width=0.44\\textwidth]{phi_KED.pdf} &\n \\includegraphics[width=0.45\\textwidth]{phidot_KED.pdf} \\\\\n \n \\end{array}\n $\n\n$\n \\begin{array}{c c}\n \\includegraphics[width=0.44\\textwidth]{HubbleParameter_KED.pdf} &\n \\includegraphics[width=0.454\\textwidth]{acceleration_KED.pdf} \\\\\n \n \\end{array}\n $\n\n$\n \\begin{array}{c c}\n \\includegraphics[width=0.44\\textwidth]{epsilon1.pdf} &\n \\includegraphics[width=0.429\\textwidth]{epsilon2.pdf} \\\\\n \n \\end{array}\n $\n\n$\n \\begin{array}{c c}\n \\includegraphics[width=0.44\\textwidth]{epsilon3.pdf} &\n \\includegraphics[width=0.463\\textwidth]{N_KED.pdf} \\\\\n \n \\end{array}\n $\n\n$\n \\begin{array}{c c}\n \\includegraphics[width=0.44\\textwidth]{a_KED.pdf} &\n \\includegraphics[width=0.43\\textwidth]{Loga_KED.pdf} \\\\\n \n \\end{array}\n $\n\n$\n \\begin{array}{c c}\n \\includegraphics[width=0.44\\textwidth]{wphi.pdf} \n \\end{array}\n $\n\n$\n \\begin{array}{c c}\n \\includegraphics[width=0.44\\textwidth]{rho_KED.pdf} &\n \\includegraphics[width=0.463\\textwidth]{rw_KED.pdf} \\\\\n \n \\end{array}\n $\n\n$\n \\begin{array}{c c}\n \\includegraphics[width=0.438\\textwidth]{Osc_phi.pdf} &\n \\includegraphics[width=0.463\\textwidth]{Osc_PhiDot.pdf} \\\\\n \n \\includegraphics[width=0.463\\textwidth]{Osc_adoubledot.pdf} &\n \\includegraphics[width=0.41\\textwidth]{Osc_e1.pdf} \\\\\n \n \\end{array}\n $\n\n $\\phi_B$ Inflation $t/t_{pl}$ $\\phi_{*}$ $N_{inf}$ \n ------------- ----------------- ---------------------------- ------------------ --------------\n 125 starts $2.19142 \\times 10^5$ 1.945 5.324\n ends $4.41278 \\times 10^6$ 0.201683 \n 175 starts $ 1.49064 \\times 10^5$ 3.91 12.1599\n ends $6.92933 \\times10^6$ 0.257044 \n 200 starts $1.29018 \\times 10^5$ 5.179 15.0501\n ends $8.15411 \\times 10^6$ 0.257289 \n 350 starts $7.1989 \\times 10^4$ 16.2 48.64516\n ends $1.54968 \\times 10^7$ 0.251808 \n 375 starts $6.7118 \\times 10^4$ 18.62 53.2791\n ends $1.66544 \\times 10^7$ 0.2607 \n 390 starts $6.4485 \\times 10^4$ 20.08 59.984\n ends $1.7398 \\times 10^7$ 0.256375 \n 395 starts $6.3645 \\times 10^4$ 20.67 61.07\n ends $1.76853 \\times 10^7$ 0.250834 \n 400 starts $6.2806 \\times 10^4$ 21.2 62.585\n ends $1.7847 \\times 10^7$ 0.260837 \n 500 starts $5.0076 \\times 10^4$ 33.19 98.475\n ends $2.26954 \\times 10^7$ 0.2596 \n 700 starts $3.5644 \\times 10^4$ 65.21 193.841\n ends $3.24313\\times 10^7$ 0.250473 \n 800 starts $3.1144 \\times 10^4$ 85.2 253.527\n ends $3.71644 \\times 10^7$ 0.2261615 \n 850 starts $2.9280 \\times 10^4$ 95.33 286.21\n ends $3.95935 \\times 10^7$ 0.2589 \n 900 starts $2.7618 \\times 10^4$ 107.6 320.829\n ends $4.1991 \\times 10^7$ 0.259767 \n 1000 starts $2.4848 \\times 10^4$ 132.7 394.839\n ends $4.67784 \\times 10^7$ 0.26261 \n\n : This table shows the values of the number of e-folds, $N_{inf}$, for various initial values $\\phi_B$ for the potential of the form $V(\\phi)= \\frac{3}{4}M^2 \\left( \\phi - M_{Pl}\\right)^2$ with $M=1.4286 \\times 10^{-6}m_{Pl}$ in the Jordan frame for KED case with $\\dot{\\phi_B}<0$.\n\n\\[Table1\\]\n\nDiscussion of the result {#DR}\n========================\n\nThis work involves exploring the background dynamics of an effective quantum corrected BD cosmology sourced by a scalar field with the Starobinsky potential in the Jordan frame. ",
"Emphasizing on the fact that, though Einstein’s frame is equivalent classically to Jordan’s frame up to a conformal transformation, it is no longer the case after quantization, we present the classical equations of motion for background dynamics in both frames. ",
"Then, we consider the quantum corrected effective equations of motion in Jordan’s frame and analyze in detail whether the bouncing conditions are satisfied when the energy density of the scalar field $\\rho_{\\phi}$ becomes maximum, that is $\\rho_{\\phi} = \\rho_c$. We find two distinct equations of motion for the Hubble parameter, namely, Eqs.(\\[H1\\]) and (\\[H2\\]). ",
"Though, the Friedmann equation together with the Klein-Gordon equation is sufficient to solve the background dynamics, we use the Raychaudhuri equation to check whether the second condition of a bounce is satisfied or not. ",
"To this effort we derive the corresponding Raycahudhuri equation for each of the Friedmann equations (\\[H1\\]) and (\\[H2\\]). ",
"Thus, analyzing the quantum corrected Raychaudhuri equation we show explicitly that for the universe given by Eq.(\\[H1\\]) one can achieve a bounce with $\\dot{\\phi}_B<0$. For the dynamics given by Eq.(\\[H2\\]) the allowed initial value is too restricted for bounce to occur. ",
"In fact, only one initial value is allowed which is the potential completely-dominated bounce. ",
"Since in this paper we are mainly interested in the initially kinetic energy dominant case, this is not considered.",
"\n\nAfter deriving the Raychaudhuri equation and analyzing the bouncing conditions, we go on to define an effective equation of state in the Jordan frame. ",
"In order to do so, we first introduce an effective pressure due to the scalar field. ",
"To obtain this, we can always assume that there exists a continuity equation, the same as in LQC of GR. ",
"Since we already have the expression of the effective energy density in terms of the scalar field and its velocity $\\rho_{\\phi}= \\frac{\\beta}{4}\\dot{\\phi}^2 + \\frac{\\phi V(\\phi)}{M_{Pl}}$, and as the scalar field must satisfy the Klein-Gordon equation, therefore, after some simple calculation it is straightforward to obtain an effective pressure.",
"\n\nNext we plot the effective equation of state $w_{\\phi}$ starting with at the bouncing point. ",
"From Fig.\\[wphi\\] we notice three phases of the evolution namely. ",
"bouncing phase where quantum geometric effects dominate, transition from bouncing to classical phase, and the slow roll inflation phase. ",
"As in the bouncing phase, the kinetic energy dominates over the potential energy, so we have $w_{\\phi} \\simeq 1$. Then, it transits from $+1$ to $-1$ in the transition phase, and afterward, the universe enters the slow roll phase of inflation when the potential energy dominates. ",
"We also note the universality of the solution in the bouncing phase from Fig.\\[wphi\\].",
"\n\nThen, we analyze the slow roll inflation. ",
"As one of the main object of this article is to see if starting with an initially bouncing phase one can land up with a slow roll phase of inflation, and if so, for what values of initial conditions we obtain 60 number of e-folds. ",
"In order to study the slow roll inflation we choose the dynamics given by Eq.(\\[H1\\]) for the Hubble parameter together with the Klein-Gordon equation. ",
"The dynamics dictated by Eq.(\\[H2\\]) is not considered as it allows only one initial condition for bounce to occur which is the extreme case of potential energy completely-dominated bounce. ",
"Also, we note that for dynamics generated by Eq.(\\[H1\\]) for any $\\phi_B$, the only allowed sign of $\\dot{\\phi}_B$ is negative as $\\dot{\\phi_B}>0$ does not satisfy the second condition of a bounce. ",
"And it is obvious that the value of $\\dot{\\phi}_B$ is fixed by $(\\rho_{\\phi})_B=\\rho_c$ for any given $\\phi_B$. This allows us to investigate this interesting situation in which one is free to displace, initially, the scalar field away from the minimum of the potential and setting its initial velocity negative and see if the scalar field rolls down the potential slowly enough to give rise to a slow roll inflation. ",
"In BD theory, there are three nontrivial parameters to determine if or not a slow roll phase of inflation is attained. ",
"In this work, we obtain the slow roll phase of inflation in quantum corrected BD theory in the Jordan frame for initial values $\\phi_{B}\\gtrapprox 100 m_{Pl}$. We plot all the relevant cosmological parameters, and show that $\\epsilon_1,~\\epsilon_2, \\epsilon_3 \\ll 1$ in the period $(10^{4} \\sim 10^7)t/t_{Pl}$. We also calculate the number of e-folds, $N_{inf}$, generated by various initial conditions at the bounce and dispaly all our results in Table \\[Table1\\]. ",
"We observe that as we increase $\\phi_B$ the number $N_{inf}$ increases and reaches its value $60$ for $\\phi_B=395~m_{Pl}$. Finally we plot $N_{inf}$ w.r.t $\\phi_B$.\n\n**Acknowledgement** A.W. is supported in part by the National Natural Science Foundation of China (NNSCF) with the Grants Nos. ",
"11375153 and 11675145. ",
"T.Z. is supported in part by NFSC with Grant No. ",
"11675143 and the Fundamental Research Funds for the Provincial Universities of Zhejiang in China with Grant No. ",
"RF-A2019015. ",
"M.S. expresses sincere gratitude towards Prof. Yongge Ma for his valuable suggestions and the fruitful discussion.",
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"\n\n[^1]: E-mail address: manabendra@zjut.edu.cn\n\n[^2]: E-mail address: zhut05@zjut.edu.cn\n\n[^3]: E-mail address: Anzhong$\\_$Wang@baylor.edu\n"
] | {
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[
"\n982 So.2d 696 (2008)\nOLIVA\nv.\nSTATE.",
"\nNo. ",
"2D07-4092.",
"\nDistrict Court of Appeal of Florida, Second District.",
"\nMay 28, 2008.",
"\nDecision without published opinion. ",
"Affirmed.",
"\n"
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"Osbern FitzOsbern\n\n__NOTOC__\n\nOsbern fitzOsbern (c. 1032–1103) was a Norman churchman.",
"\n\nBiography\nOsbern was a relative of King Edward the Confessor as well as being a royal chaplain. ",
"During Edward's reign he received the church at Bosham, near Chichester. ",
"He was one of those present at the consecration of Westminster Abbey at Christmas 1065. ",
"He was a steward for King William I of England during his reign, as well as being a friend of the king. ",
"The story that he became William's chancellor is based entirely on a charter that modern historians have declared mostly spurious. ",
"He became Bishop of Exeter in 1072, and was consecrated at St. Paul's in London on 27 May 1072 by the Archbishop of Canterbury, Lanfranc.",
"\n\nOsbern was present at the church councils held in 1072 and 1075. ",
"Osbern was present at the first Christmas court held by King William II of England after his accession. ",
"Osbern did not attend the church council held by Anselm, the new Archbishop of Canterbury in 1102, as he was ill. ",
"He became embroiled in a dispute with the monks of Battle Abbey, who had established a priory in Exeter. ",
"The cathedral chapter of Exeter objected to the priory establishing a graveyard or ringing their bells, and both sides appealed to Anselm, who ruled in Battle's favor on the bell issue. ",
"The dispute over the graveyard was still ongoing in 1102, when Pope Paschal II wrote to Osbern ordering him to allow the priory to establish a graveyard for their benefactors.",
"\n\nOsbern FitzOsbern died in 1103, having gone blind before his death. ",
"William fitz Osbern, Earl of Hereford was his brother. ",
"Their father was Osbern de Crépon, a guardian and seneschal to the young Duke William. ",
"Frank Barlow, a medieval historian, described Osbern as \"unsociable\".",
"\n\nCitations\n\nSources\n\nExternal links\n \"Entry for Osbern\" in George Oliver's Lives of the Bishops of Exeter\n\nCategory:1103 deaths\nCategory:11th-century Roman Catholic bishops\nCategory:Normans in England\nCategory:Bishops of Exeter\nCategory:Year of birth uncertain\nCategory:Devon Domesday Book tenants-in-chief\nCategory:Norman clerics given benefices in England"
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[
". ",
"Let x be o(-50). ",
"Calculate r(x).",
"\n1\nLet m(s) = 5 - 1 - 2*s - 7 + 2. ",
"Let l be (-10)/(-15)*(-3)/2*1. ",
"Calculate m(l).",
"\n1\nLet a(s) be the first derivative of 1/3*s**3 - 152 - 23*s - 3/2*s**2. ",
"Calculate a(6).",
"\n-5\nLet m(h) = 139*h - 48. ",
"Let r(j) = -250*j + 88. ",
"Let t(n) = 9*m(n) + 5*r(n). ",
"Suppose 5*i = -3*u + 43, 0 = -5*i + 16 + 9. ",
"Give t(u).",
"\n14\nSuppose -8 - 28 = -6*y. ",
"Let m(h) be the first derivative of -h**2/2 - 5*h - 2071. ",
"Give m(y).",
"\n-11\nSuppose -32*a + 16 = -28*a. ",
"Suppose -4*p - 3*v + 76 = -0*v, -a = -v. ",
"Let d(l) = 5 - p + 8 + l. Determine d(-6).",
"\n-9\nLet k(j) = 21*j. ",
"Let o(r) = -r**2 + 2*r + 63. ",
"Let p be o(-7). ",
"Suppose 17 = 5*u - 3*m - 0*m, p = u - m - 5. ",
"Calculate k(u).",
"\n21\nLet f(q) be the first derivative of 0*q + 1/24*q**4 - 15 + 1/3*q**3 + 9*q**2. ",
"Let h(b) be the second derivative of f(b). ",
"What is h(-3)?",
"\n-1\nLet u = -91 - -81. ",
"Let d(n) be the second derivative of -n**4/12 - 5*n**3/3 - n**2 - 17*n + 1. ",
"Determine d(u).",
"\n-2\nLet i(x) = 2*x**3 - 2*x**2 + 3*x + 3. ",
"Suppose 1068*s = 1045*s + 46. ",
"Determine i(s).",
"\n17\nLet p(x) = x**3 - x + 2. ",
"Suppose -35*b + 22*b + 39 = 0. ",
"Suppose 36*k - b*k - 99 = 0. ",
"Give p(k).",
"\n26\nSuppose 2*r - 3924 = 4*b - 5*b, -3*b = -3*r - 11745. ",
"Let o(s) = -b*s**2 + 3918*s**2 + s**3 - s - 5. ",
"Give o(0).",
"\n-5\nLet u(g) = 64*g + 62. ",
"Let x(m) = -41*m - 41. ",
"Let n(o) = 5*u(o) + 8*x(o). ",
"Calculate n(-9).",
"\n54\nLet f(u) = 53*u**3 - 17*u - 41*u**3 - 15*u**2 + 19 - 11*u**3. ",
"What is f(16)?",
"\n3\nLet w be -76 + 7/((-7)/(-4)). ",
"Let m be 16/w - 22/(-18). ",
"Let c(i) = -52*i + 101*i - 6 + 5 - 55*i. ",
"Determine c(m).",
"\n-7\nLet k(v) = -v**2 - 27*v + 64. ",
"Let n = 7933 + -7963. ",
"What is k(n)?",
"\n-26\nSuppose -6452*o + 70 = -6382*o. ",
"Let m(x) = -21*x**3 - 3*x**2 + 3*x + 1. ",
"What is m(o)?",
"\n-20\nLet s(i) be the third derivative of 0 + 4*i + 15*i**2 - 2*i**3 - 1/120*i**6 - 2/15*i**5 - 1/6*i**4. ",
"What is s(-8)?",
"\n20\nLet a(g) = 7*g**3 + 30*g**2 - 5*g + 53. ",
"Let y(f) = 20*f**3 + 90*f**2 - 14*f + 153. ",
"Let r(v) = -17*a(v) + 6*y(v). ",
"Calculate r(-30).",
"\n-13\nLet z(o) = o**2 - 14*o - 14. ",
"Suppose -23*t - 4*t = -270. ",
"Calculate z(t).",
"\n-54\nLet b(c) = -1826*c + 1822*c + 136 - 89. ",
"What is b(15)?",
"\n-13\nLet x(n) = -17*n + 61. ",
"Let m(y) = y + 20. ",
"Let r(i) = 3*m(i) - x(i). ",
"What is r(-4)?",
"\n-81\nLet z(r) = -7*r + 6. ",
"Let q be z(-2). ",
"Let f = 21 - q. Let s(h) = 3*h - 4 + 2 + 4 - f. Give s(-2).",
"\n-5\nLet a(k) = -k**3 - 4*k**2 + 5*k + 1. ",
"Let g(u) = 2*u - 37. ",
"Let s be g(21). ",
"Suppose p = -s*t + 6 + 7, 1 = -3*p + 5*t. ",
"Let q be (30/p)/(-6 - -4). ",
"Calculate a(q).",
"\n1\nLet k(r) = 13*r - 35. ",
"Let a(i) = -6*i + 17. ",
"Let h(u) = 9*a(u) + 4*k(u). ",
"Calculate h(6).",
"\n1\nLet c(u) = 4*u**3 - 3*u**2 + 2*u + 2. ",
"Let s = 65 - 62. ",
"Let l be ((-21)/(-6))/(2 - s/2). ",
"Suppose -5*a + 29 = 2*t, 3*t + t - 3*a + l = 0. ",
"What is c(t)?",
"\n26\nLet d(x) be the second derivative of 5/6*x**3 + 1/12*x**4 - 7/2*x**2 + 0 - x. Suppose -173*c = -4*g - 169*c - 40, 10 = -5*g - 3*c. ",
"Determine d(g).",
"\n-7\nLet l(b) = -b - 2. ",
"Suppose 0 = -5*w - 67 + 767. ",
"Let d = 134 - w. Give l(d).",
"\n4\nLet r(h) be the first derivative of -h**4/4 + 22*h**3/3 + h**2/2 + 38*h + 4556. ",
"What is r(22)?",
"\n60\nSuppose 0 = 4*i - 4*k - 536, 6*k - 4*k = 4*i - 540. ",
"Let u = -130 + i. Let l(t) = 2 + t - t + 7*t**2 - t**3 - 4 - 3*t. ",
"Calculate l(u).",
"\n16\nLet u be (1 + 3/(-12))*8. ",
"Let z(l) = -4*l + 4503 + 0*l - 4501 + l**2. ",
"Calculate z(u).",
"\n14\nLet m = -17393 + 17396. ",
"Let s(v) be the third derivative of -4*v**2 - 1/12*v**4 + 0 + 1/3*v**m + 0*v. ",
"Determine s(5).",
"\n-8\nLet q(g) = 3*g - 17. ",
"Suppose 269*k - 166*k = -1236. ",
"Calculate q(k).",
"\n-53\nLet h(p) be the third derivative of -p**8/20160 - p**7/630 - 97*p**5/60 - p**3/6 + 2*p**2 - 54*p. ",
"Let g(t) be the third derivative of h(t). ",
"What is g(-9)?",
"\n-9\nSuppose -5*y + 189 = 3*x, 59 + 4 = x - 2*y. ",
"Let a = -56 + x. Let k(c) = 2*c**2 - c + 21. ",
"Let z(d) = -d**2 + d - 11. ",
"Let r(p) = a*z(p) + 4*k(p). ",
"Give r(-5).",
"\n17\nLet u(l) = -2*l**3 + l. Let j(h) = -4*h**3 - 3*h - 1. ",
"Let o(i) = -j(i) - 2*u(i). ",
"Let r = 55 - 56. ",
"Calculate o(r).",
"\n-8\nLet k(x) be the second derivative of 2*x**4 - x**3/3 + x**2/2 - 2549*x. ",
"Calculate k(-1).",
"\n27\nLet l(b) = -14*b + 4. ",
"Suppose 18*y = 2863 - 2917. ",
"Calculate l(y).",
"\n46\nLet a be ((-1)/7)/((-82)/574). ",
"Let r(i) = -i + 0*i + 0 - 5*i + a. Determine r(4).",
"\n-23\nLet w be ((-1344)/56)/(3/2). ",
"Let x(q) = q**2 + 14*q - 12. ",
"Calculate x(w).",
"\n20\nLet h(y) = -y - 1. ",
"Let l(p) = p**2 + 25*p - 35. ",
"Let k be l(-28). ",
"Suppose 136 = 32*o - k*o. ",
"Give h(o).",
"\n7\nLet x(k) = -23*k - 236. ",
"Suppose 70*b + 977 = -23*b - 46. ",
"Calculate x(b).",
"\n17\nLet h be (-2911)/355 - (1/5)/(-1). ",
"Let c(z) = -z**3 - 7*z**2 + 7*z - 5. ",
"What is c(h)?",
"\n3\nLet d(y) = y**2 - 24*y + 29*y + 555 - 534. ",
"Calculate d(-8).",
"\n45\nLet h(q) = 59 - 28*q**3 - 42*q**2 + 27*q**3 + 19711*q - 19667*q. ",
"What is h(-43)?",
"\n16\nLet b(i) = i**2 - 2*i - 5. ",
"Let v be (24/(-28))/((-1)/791). ",
"Let t = -682 + v. What is b(t)?",
"\n19\nSuppose -31 = 3*w - 28. ",
"Let l(b) = 4*b**2 + 7*b + 9. ",
"What is l(w)?",
"\n6\nLet y(b) be the third derivative of 0*b**4 + 0*b - 1/120*b**6 - 10*b**2 + 0 + 1/20*b**5 - 1/6*b**3. ",
"Give y(4).",
"\n-17\nLet w(t) = 3*t - 2. ",
"Let i(j) = -1. ",
"Let k(d) = -3*i(d) + w(d). ",
"Suppose f = 4*n - 26, -4*n - 59 = -5*f - 93. ",
"What is k(f)?",
"\n-5\nLet p = 83 - 4. ",
"Let n = 4 + -2. ",
"Let w be n/(-11) + p/11 + -2. ",
"Let c(t) = t - 6. ",
"Give c(w).",
"\n-1\nLet z(a) = -32*a + 237. ",
"Suppose 36 = 4*o, -4*b + 68 - 9 = 3*o. ",
"Calculate z(b).",
"\n-19\nLet n(g) = 2*g**2 + 35*g + 49. ",
"Suppose -49*y + 312 + 600 = -106*y. ",
"Give n(y).",
"\n1\nLet x(q) = -22*q - 43 + 9*q + 36 - 4*q**2. ",
"What is x(-5)?",
"\n-42\nLet s(n) = -n + 29. ",
"Let i(a) = -16. ",
"Let b(d) = -9*i(d) - 5*s(d). ",
"What is b(6)?",
"\n29\nLet h(x) = -x**2 - 9*x - 7. ",
"Let m be 507/27 + 4/18. ",
"Let l = 53 - m. Suppose -7*z + 2*z - l = j, z - 5*j = 14. ",
"What is h(z)?",
"\n11\nLet f(q) = q**2 + 13*q + 14. ",
"Let b be f(-11). ",
"Let p be 42/8 - 6/b. Let j(n) = 2*n + 2. ",
"What is j(p)?",
"\n14\nLet a(y) = -15*y**2 - y**3 - 14*y + 21 - 2*y**2 + y**2. ",
"Suppose -3*h = -2*s + 9, -183*s + 180*s = 5*h + 110. ",
"What is a(s)?",
"\n6\nLet g = -453 - 455. ",
"Let r = 903 + g. Let i(p) = p**2 + 8*p + 12. ",
"Calculate i(r).",
"\n-3\nLet s be (-34)/6 - 2/(-3). ",
"Suppose -2*f + 73 = w, 0 = 2*w - 4*f + 6*f - 156. ",
"Let d(u) = -83*u**2 - w*u**2 + 165*u**2 - 2*u. ",
"Calculate d(s).",
"\n-15\nSuppose 3*v - 14*v + 22 = 0. ",
"Let a(x) = -2*x - 5*x**2 - 17*x + 7*x + x**2 + x**3 - 20 - 9*x**v. ",
"What is a(14)?",
"\n8\nLet b(q) = -5*q - 18 + 30 - 8*q + 3*q**2 - q**2 + 27. ",
"Determine b(5).",
"\n24\nLet c(l) = 10*l**2 + 13*l - 12. ",
"Let r = 8611 - 8610. ",
"Calculate c(r).",
"\n11\nLet d(x) = -x - 30. ",
"Let c be d(-14). ",
"Let h be (-58)/c + -1 + 55/40. ",
"Let w(f) = -4*f + 6. ",
"Determine w(h).",
"\n-10\nLet s(r) be the first derivative of r**4/4 - 8*r**3/3 + 3*r + 2. ",
"Let o(z) = 9 + 33*z + 25*z - 57*z - 3 - 1. ",
"Let t be o(3). ",
"Give s(t).",
"\n3\nSuppose -31*m + 6*m + 100 = 0. ",
"Suppose 2*o = 5*a + 2, -2*a - 4 = -0*o - m*o. ",
"Let c(w) = 4*w**3 - w**2 + w. Give c(o).",
"\n4\nLet w(y) = 8*y**2 - 3*y + 5. ",
"Let p(x) = x**2 - x + 1. ",
"Let j(v) = -6*p(v) + w(v). ",
"Suppose 761*n + 428 = 654*n. ",
"Calculate j(n).",
"\n19\nLet d = 82750 - 82753. ",
"Let g(p) = 5*p**3 - 10*p**2 - 6*p + 1. ",
"Let c(w) = -9*w**3 + 19*w**2 + 12*w - 1. ",
"Let i(y) = -6*c(y) - 11*g(y). ",
"Give i(d).",
"\n4\nSuppose 10 = -2*p - 4. ",
"Let d(o) = 23*o**3 + 18*o**2 - 16*o + 5. ",
"Let k(v) = 61*v**3 + 46*v**2 - 40*v + 14. ",
"Let i(x) = 8*d(x) - 3*k(x). ",
"Calculate i(p).",
"\n5\nLet w(z) = -20*z - 17. ",
"Suppose 0 = 336*c - 339*c + 51. ",
"Let l be 17/3 - (-1)/3. ",
"Let q(m) = 7*m + 6. ",
"Let x(t) = c*q(t) + l*w(t). ",
"Give x(-5).",
"\n5\nLet k(p) = -2*p**2 - p + 9. ",
"Suppose -74 = 50*h + 126. ",
"Let i(x) = 3*x**2 + x - 12. ",
"Let b(l) = h*k(l) - 3*i(l). ",
"Calculate b(3).",
"\n-6\nLet g(b) be the second derivative of b**4/12 - 2*b**3 + 31*b**2/2 - 71*b - 10. ",
"Give g(5).",
"\n-4\nLet x(u) = 8*u**2 - 37*u - 11. ",
"Let l(c) = -8*c**2 + 37*c + 12. ",
"Let q(k) = 2*l(k) + 3*x(k). ",
"What is q(5)?",
"\n6\nLet w(c) = 9*c - 179. ",
"Let r = -1663 + 1678. ",
"What is w(r)?",
"\n-44\nLet w(m) = -128 + 121 - m**3 + 4*m**2 + 0*m**2 + 7*m. ",
"Let v be w(5). ",
"Let i(j) = 36*j**2 + 6*j - 37*j**2 - v + 2*j. ",
"Give i(6).",
"\n9\nLet k(n) be the first derivative of n**5/20 - 2*n**4/3 - 4*n**3/3 - 11*n**2/2 - 52*n + 19. ",
"Let l(r) be the first derivative of k(r). ",
"Calculate l(9).",
"\n-2\nSuppose -8*i + 4 = -36. ",
"Suppose 5*s + 3*v + 11 = 0, 4*s + i*v + 5 - 4 = 0. ",
"Let c(t) = -t**2 - 5*t - 5. ",
"Determine c(s).",
"\n-1\nLet x be (-30)/(-50) - 8/5 - -3. ",
"Suppose -5*t = -x*t - 6. ",
"Let s(m) = 2 - 5*m + 125*m**2 + m - 124*m**t. ",
"Calculate s(5).",
"\n7\nLet j be 3/(-18)*-4*372/(-62). ",
"Let i(g) = 1. ",
"Let k(m) = -m - 4. ",
"Let a(q) = q**2 + 8*q - 2. ",
"Let t be a(-8). ",
"Let v(w) = t*i(w)"
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[
"Q:\n\nPartially transposing a data frame\n\nI have a data frame with data like this:\nA B C D\na1 b1 c1 d1\na1 b1 c2 d2\na1 b1 c3 d3\na2 b2 c1 d1\na2 b2 c3 d3\n\nHow would I go about transforming that into?",
"\nA B c1 c2 c3\na1 b1 d1 d2 d3\na2 b2 d1 d3\n\nA:\n\nIn base R, you can use reshape():\nreshape(mydf, direction = \"wide\", idvar = c(\"A\", \"B\"), timevar = \"C\")\n# A B D.c1 D.c2 D.c3\n# 1 a1 b1 d1 d2 d3\n# 4 a2 b2 d1 <NA> d3\n\nYou can also use tidyr and dplyr together, like this:\nlibrary(dplyr)\n# devtools::install_github(\"hadley/tidyr\")\nlibrary(tidyr)\nmydf %>% group_by(A, B) %>% spread(C, D)\n# Source: local data frame [2 x 5]\n# \n# A B c1 c2 c3\n# 1 a1 b1 d1 d2 d3\n# 2 a2 b2 d1 NA d3\n\nA:\n\nThis is a good place to use the reshape2 library. ",
"You can just do\nlibrary(reshape2)\ndcast(dd, A+B~C)\n\nto get\n A B c1 c2 c3\n1 a1 b1 d1 d2 d3\n2 a2 b2 d1 <NA> d3\n\nas desired.",
"\n\n"
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[
"/*\n * Copyright (c) 2017-2020 Arm Limited.",
"\n *\n * SPDX-License-Identifier: MIT\n *\n * Permission is hereby granted, free of charge, to any person obtaining a copy\n * of this software and associated documentation files (the \"Software\"), to\n * deal in the Software without restriction, including without limitation the\n * rights to use, copy, modify, merge, publish, distribute, sublicense, and/or\n * sell copies of the Software, and to permit persons to whom the Software is\n * furnished to do so, subject to the following conditions:\n *\n * The above copyright notice and this permission notice shall be included in all\n * copies or substantial portions of the Software.",
"\n *\n * THE SOFTWARE IS PROVIDED \"AS IS\", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR\n * IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY,\n * FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. ",
"IN NO EVENT SHALL THE\n * AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER\n * LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM,\n * OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE\n * SOFTWARE.",
"\n */\n#include \"arm_compute/graph.h\"\n#ifdef ARM_COMPUTE_CL\n#include \"arm_compute/runtime/CL/Utils.h\"\n#endif /* ARM_COMPUTE_CL */\n#include \"support/ToolchainSupport.h\"\n#include \"utils/CommonGraphOptions.h\"\n#include \"utils/GraphUtils.h\"\n#include \"utils/Utils.h\"\n\nusing namespace arm_compute;\nusing namespace arm_compute::utils;\nusing namespace arm_compute::graph::frontend;\nusing namespace arm_compute::graph_utils;\n\n/** Example demonstrating how to implement AlexNet's network using the Compute Library's graph API */\nclass GraphAlexnetExample : public Example\n{\npublic:\n GraphAlexnetExample()\n : cmd_parser(), common_opts(cmd_parser), common_params(), graph(0, \"AlexNet\")\n {\n }\n bool do_setup(int argc, char **argv) override\n {\n // Parse arguments\n cmd_parser.parse(argc, argv);\n cmd_parser.validate();\n\n // Consume common parameters\n common_params = consume_common_graph_parameters(common_opts);\n\n // Return when help menu is requested\n if(common_params.help)\n {\n cmd_parser.print_help(argv[0]);\n return false;\n }\n\n // Checks\n ARM_COMPUTE_EXIT_ON_MSG(arm_compute::is_data_type_quantized_asymmetric(common_params.data_type), \"QASYMM8 not supported for this graph\");\n\n // Print parameter values\n std::cout << common_params << std::endl;\n\n // Get trainable parameters data path\n std::string data_path = common_params.data_path;\n\n // Create a preprocessor object\n const std::array<float, 3> mean_rgb{ { 122.68f, 116.67f, 104.01f } };\n std::unique_ptr<IPreprocessor> preprocessor = arm_compute::support::cpp14::make_unique<CaffePreproccessor>(mean_rgb);\n\n // Create input descriptor\n const auto operation_layout = common_params.data_layout;\n const TensorShape tensor_shape = permute_shape(TensorShape(227U, 227U, 3U, 1U), DataLayout::NCHW, operation_layout);\n TensorDescriptor input_descriptor = TensorDescriptor(tensor_shape, common_params.data_type).set_layout(operation_layout);\n\n // Set weights trained layout\n const DataLayout weights_layout = DataLayout::NCHW;\n\n graph << common_params.target\n << common_params.fast_math_hint\n << InputLayer(input_descriptor, get_input_accessor(common_params, std::move(preprocessor)))\n // Layer 1\n << ConvolutionLayer(\n 11U, 11U, 96U,\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/conv1_w.npy\", weights_layout),\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/conv1_b.npy\"),\n PadStrideInfo(4, 4, 0, 0))\n .set_name(\"conv1\")\n << ActivationLayer(ActivationLayerInfo(ActivationLayerInfo::ActivationFunction::RELU)).set_name(\"relu1\")\n << NormalizationLayer(NormalizationLayerInfo(NormType::CROSS_MAP, 5, 0.0001f, 0.75f)).set_name(\"norm1\")\n << PoolingLayer(PoolingLayerInfo(PoolingType::MAX, 3, operation_layout, PadStrideInfo(2, 2, 0, 0))).set_name(\"pool1\")\n // Layer 2\n << ConvolutionLayer(\n 5U, 5U, 256U,\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/conv2_w.npy\", weights_layout),\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/conv2_b.npy\"),\n PadStrideInfo(1, 1, 2, 2), 2)\n .set_name(\"conv2\")\n << ActivationLayer(ActivationLayerInfo(ActivationLayerInfo::ActivationFunction::RELU)).set_name(\"relu2\")\n << NormalizationLayer(NormalizationLayerInfo(NormType::CROSS_MAP, 5, 0.0001f, 0.75f)).set_name(\"norm2\")\n << PoolingLayer(PoolingLayerInfo(PoolingType::MAX, 3, operation_layout, PadStrideInfo(2, 2, 0, 0))).set_name(\"pool2\")\n // Layer 3\n << ConvolutionLayer(\n 3U, 3U, 384U,\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/conv3_w.npy\", weights_layout),\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/conv3_b.npy\"),\n PadStrideInfo(1, 1, 1, 1))\n .set_name(\"conv3\")\n << ActivationLayer(ActivationLayerInfo(ActivationLayerInfo::ActivationFunction::RELU)).set_name(\"relu3\")\n // Layer 4\n << ConvolutionLayer(\n 3U, 3U, 384U,\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/conv4_w.npy\", weights_layout),\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/conv4_b.npy\"),\n PadStrideInfo(1, 1, 1, 1), 2)\n .set_name(\"conv4\")\n << ActivationLayer(ActivationLayerInfo(ActivationLayerInfo::ActivationFunction::RELU)).set_name(\"relu4\")\n // Layer 5\n << ConvolutionLayer(\n 3U, 3U, 256U,\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/conv5_w.npy\", weights_layout),\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/conv5_b.npy\"),\n PadStrideInfo(1, 1, 1, 1), 2)\n .set_name(\"conv5\")\n << ActivationLayer(ActivationLayerInfo(ActivationLayerInfo::ActivationFunction::RELU)).set_name(\"relu5\")\n << PoolingLayer(PoolingLayerInfo(PoolingType::MAX, 3, operation_layout, PadStrideInfo(2, 2, 0, 0))).set_name(\"pool5\")\n // Layer 6\n << FullyConnectedLayer(\n 4096U,\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/fc6_w.npy\", weights_layout),\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/fc6_b.npy\"))\n .set_name(\"fc6\")\n << ActivationLayer(ActivationLayerInfo(ActivationLayerInfo::ActivationFunction::RELU)).set_name(\"relu6\")\n // Layer 7\n << FullyConnectedLayer(\n 4096U,\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/fc7_w.npy\", weights_layout),\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/fc7_b.npy\"))\n .set_name(\"fc7\")\n << ActivationLayer(ActivationLayerInfo(ActivationLayerInfo::ActivationFunction::RELU)).set_name(\"relu7\")\n // Layer 8\n << FullyConnectedLayer(\n 1000U,\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/fc8_w.npy\", weights_layout),\n get_weights_accessor(data_path, \"/cnn_data/alexnet_model/fc8_b.npy\"))\n .set_name(\"fc8\")\n // Softmax\n << SoftmaxLayer().set_name(\"prob\")\n << OutputLayer(get_output_accessor(common_params, 5));\n\n // Finalize graph\n GraphConfig config;\n\n config.num_threads = common_params.threads;\n config.use_tuner = common_params.enable_tuner;\n config.tuner_mode = common_params.tuner_mode;\n config.tuner_file = common_params.tuner_file;\n\n // Load the precompiled kernels from a file into the kernel library, in this way the next time they are needed\n // compilation won't be required.",
"\n if(common_params.enable_cl_cache)\n {\n#ifdef ARM_COMPUTE_CL\n restore_program_cache_from_file();\n#endif /* ARM_COMPUTE_CL */\n }\n\n graph.finalize(common_params.target, config);\n\n // Save the opencl kernels to a file\n if(common_opts.enable_cl_cache)\n {\n#ifdef ARM_COMPUTE_CL\n save_program_cache_to_file();\n#endif /* ARM_COMPUTE_CL */\n }\n\n return true;\n }\n void do_run() override\n {\n // Run graph\n graph.run();\n }\n\nprivate:\n CommandLineParser cmd_parser;\n CommonGraphOptions common_opts;\n CommonGraphParams common_params;\n Stream graph;\n};\n\n/** Main program for AlexNet\n *\n * Model is based on:\n * https://papers.nips.cc/paper/4824-imagenet-classification-with-deep-convolutional-neural-networks\n * \"ImageNet Classification with Deep Convolutional Neural Networks\"\n * Alex Krizhevsky and Sutskever, Ilya and Hinton, Geoffrey E\n *\n * Provenance: https://github.com/BVLC/caffe/tree/master/models/bvlc_alexnet\n *\n * @note To list all the possible arguments execute the binary appended with the --help option\n *\n * @param[in] argc Number of arguments\n * @param[in] argv Arguments\n *\n * @return Return code\n */\nint main(int argc, char **argv)\n{\n return arm_compute::utils::run_example<GraphAlexnetExample>(argc, argv);\n}\n"
] | {
"pile_set_name": "Github"
} | [
0,
0.0096,
0.004651162790697674,
0.003703703703703704,
0.0030581039755351682,
0.01023391812865497
] | 0.005208 | 5 |
[
"From practice in a car park and over a Persian carpet to skating in one of the most prestigious hockey academies: Eurohockey meets Guila Mohajer: facing many difficulties, the first Iranian women hockey player is now studying coaching in Vierumaki Sport ... Read more»"
] | {
"pile_set_name": "Pile-CC"
} | [
0.0037313432835820895
] | 0.003731 | 5 |
[
"At D'Agosto & Howe, LLC, we are dedicated to providing workplace injury victims with clear and effective legal guidance. ",
"Whether you need help with Connecticut workers' compensation benefits or personal injury claims in general, we will work tirelessly to meet...more"
] | {
"pile_set_name": "Pile-CC"
} | [
0.01652892561983471,
0
] | 0.008264 | 5 |
[
"Laser Cut Record\n\nNot satisfyied with creating the world's first 3D printed record, Amanda Ghassaei has now created wooden, laser cut records. ",
"The results are not only striking in a visual sense, but the audible versions of the songs cut into the wood in this way are strangely haunting. ",
"Tracks that Amanda has chosen so far include Radiohead's Idioteque and The Velvet Underground's Sunday Morning.",
"\n\nHave a watch of the video to get a better sense of what the sound's actually like and to get a glimpse of the process.",
"\n\nComments: 0\n\nLeave a Comment\n\nPlease feel free to comment on this post. ",
"Please note that all comments are subject to approval and anything deemed offensive will not be published. ",
"Your email is never published nor shared. ",
"Required fields are marked *"
] | {
"pile_set_name": "Pile-CC"
} | [
0.006993006993006993,
0,
0.02702702702702703,
0,
0,
0,
0,
0
] | 0.004253 | 5 |
[
"Association of virulence factors, phylogenetic groups and antimicrobial resistance markers in Escherichia coli from Badin city, Pakistan.",
"\nEscherichia coli, the most frequent cause of UTIs has extensive genetic substructure and can be assigned to eight phylogroups, A, B1, B2, C, D, E, F and Escherichia cryptic clade I. We investigated the distribution of virulence determinants and antimicrobial resistance genes in relation to phylogenetic groups. ",
"A total of 77 E. coli isolates were collected from Civil Hospital Badin, Pakistan. ",
"Isolates were assigned phylogroups using quadruplex PCR method, while virulence and antibiotic resistance genes, blaCTX-M and blaNDM-1 were also detected using PCR. ",
"Thirty-four isolates were assigned to group B2, while 23, 2, 1, 7 and 10 isolates were assigned to F, B1, A/C, clade I/II and negative, respectively. ",
"Among virulence genes, prevalence of papC (83%) was highest followed by aer (57%), papGII (16%), papGIII (14%), cnf (9%), hly (5%) and sfa (6%). ",
"Of these isolates, 23% and 9% were positive for blaCTX-M and blaNDM-1, respectively."
] | {
"pile_set_name": "PubMed Abstracts"
} | [
0.0072992700729927005,
0.009584664536741214,
0,
0.01818181818181818,
0,
0.013793103448275862,
0.011904761904761904
] | 0.008681 | 5 |
[
"Low pulse energy Nd:YAG laser irradiation exerts a biostimulative effect on different cells of the oral microenvironment: \"an in vitro study\".",
"\nDental lasers represent a promising therapeutic tool in the treatment of periodontal and peri-implant diseases. ",
"However, their clinical application remains still limited. ",
"Here, we investigated the potential biostimulatory effect of low pulse energy neodymium:yttrium-aluminum-garnet (Nd:YAG) laser irradiation on different cells representative of the oral microenvironment and elucidated the underlying molecular mechanisms. ",
"Saos-2 osteoblasts, H-end endothelial cells, and NIH/3T3 fibroblasts pre-treated or not with photosensitizing dye methylene blue (MB), were irradiated with low pulse energy (20 mJ) and high repetition rate (50-70 Hz) Nd:YAG laser, and evaluated for cell viability and proliferation as well as for the expression of specific differentiation markers by confocal immunofluorescence and real-time RT-PCR. ",
"Changes in intracellular Ca(2+) levels after laser exposure were also evaluated in living osteoblasts. ",
"Nd:YAG laser irradiation did not affect cell viability in all the tested cell types, even when combined with pre-treatment with MB, and efficiently stimulated cell growth in the non-sensitized osteoblasts. ",
"Moreover, a significant induction in the expression of osteopontin, ALP, and Runx2 in osteoblasts, type I collagen in fibroblasts, and vinculin in endothelial cells could be observed in the irradiated cells. ",
"Pre-treatment with MB negatively affected cell differentiation in the unstimulated and laser-stimulated cells. ",
"Notably, laser irradiation also caused an increase in the intracellular Ca(2+) in osteoblasts through the activation of TRPC1 ion channels. ",
"Moreover, the pharmacologic or genetic inhibition of these channels strongly attenuated laser-induced osteopontin expression, suggesting a role for the laser-mediated Ca(2+) influx in regulating osteoblast differentiation. ",
"Low pulse energy and high repetition rate Nd:YAG laser irradiation may exert a biostimulative effect on different cells representative of the oral microenvironment, particularly osteoblasts. ",
"Pre-treatment with MB prior to irradiation hampers this effect and limits the potential clinical application of photosensitizing dyes in dental practice."
] | {
"pile_set_name": "PubMed Abstracts"
} | [
0,
0,
0,
0,
0.004987531172069825,
0,
0.0048543689320388345,
0.009615384615384616,
0.009009009009009009,
0.007142857142857143,
0,
0,
0.006535947712418301
] | 0.003242 | 5 |
[
"\nShow HN: Member Decentralized Reddit on Bitcoin (BCH) Blockchain - FreeTrade\nhttps://memberapp.github.io/#topic?type=all&start=0&limit=25&topicname=Member\n======\nFreeTrade\nIf you want to play around with it -\n\nClick 'Login' and click 'Create New Account' \\- post your address as a reply\nto this post and I'll send you some coins to get you started.",
"\n\n"
] | {
"pile_set_name": "HackerNews"
} | [
0.011461318051575931,
0
] | 0.005731 | 5 |
[
"\"Help me.\" \"",
"Help me.\" \"",
"Help me.\" \"",
"My cake is ready.\" \"",
"Time for the show.\" \"",
"Everybody on stage.\" \"",
"Ruby?\" \"",
"Is that you?\" \"",
"Goddamn it.\" \"",
"Ruby!\" \"",
"Ruby?\" \"",
"Ruby!\" \"",
"Ruby?\" \"",
"If that's you, Jupiter I got some buckshot for you, you hear me?\" \"",
"I told you, it's over!\" \"",
"I'm out, you hear me?\" \"",
"You're on your own now! ",
"I'm sorry, I can't do this anymore.\" \"",
"Hello!\" \"",
"Hey!\" \"",
"It's hot out here.\" \"",
"Stay in the car, out of the wind.\" \"",
"Well, honey, I have to stretch my legs.\" \"",
"Where the hell is everybody?\" \"",
"Hello?\" \"",
"Hey.\" \" ",
"Get up, jackass.\" \" ",
"Bobby.\" \" ",
"Fill her up, check the fluids, my friend.\" \" ",
"Oh, yeah.\" \"",
"Right away, sir.\" \" ",
"Beauty, stop it.\" \"",
"No.\" \" ",
"Piece of shit.\" \"",
"Damn it.\" \"",
"You Okay?\" \"",
"Tell me again why we couldn't fly like normal people.\" \"",
"Right, they wouldn't let your dad drive the plane.\" \"",
"Doug, relax.\" \"",
"It's their silver anniversary, and they're so happy you came.\" \" ",
"Your parents can't stand me.\" \" ",
"No, you're wrong.\" \"",
"They love you.\" \"",
"I could have taken them on a cruise.\" \"",
"You know what?\" \"",
"Stop.\" \"",
"I never ask you for anything, okay?\" \"",
"Just think how happy you'll be when Catherine comes along on our 25th.\" \"",
"It's okay.\" \"",
"It's not gonna be in a fucking trailer home.\" \"",
"Don't see too many travelers around here.\" \"",
"Where y'all headed?\" \" ",
"San Diego.\" \" ",
"California?\" \"",
"Sure didn't take the fastest road.\" \"",
"Oh, no.\" \"",
"We absolutely had to see the desert.\" \" ",
"Didn't we, Bob?\" \" ",
"Absolutely.\" \"",
"There ain't nothing to see in the desert.\" \"",
"Really?\" \"",
"Do you hear that, honey?\" \"",
"Yeah.\" \"",
"Don't you know it's illegal to smoke around a gas pump?\" \"",
"Yeah.\" \"",
"Come here.\" \"",
"Where's your mom?\" \"",
"He's a good little piggy.\" \"",
"Little baby piggy.\" \"",
"Little baby piggy.\" \"",
"Hey, Bobby, come over here.\" \" ",
"Look at this.\" \" ",
"Oh, my God.\" \"",
"That's so random.\" \"",
"You two make a perfect pair.\" \"",
"You're sweet.\" \"",
"Yeah.\" \"",
"What are you doing?\" \"",
"Don't pee there.\" \"",
"What the hell do you care?\" \"",
"Just turn around for a second.\" \"",
"Hello?\" \"",
"Don't you see that?\" \"",
"Excuse me, you have a phone I can use?\" \"",
"I can't get a signal.\" \"",
"No,ldonT\" \"Hey, Bukowski.\" \"",
"Give the cell phone a rest.\" \"",
"Sorry, Bob.\" \"",
"Duty calls.\" \"",
"Honey, you've been on the phone since we left.\" \"",
"Really.\" \"",
"I've been gone.\" \"",
"I have to make sure my store's not burning to the ground.\" \"",
"Oh, God.\" \"",
"Thanks, Brenda.\" \"",
"What the hell, Brenda?\" \"!\" \"",
"Damn it.\" \" ",
"Damn it, got it all over my hand.\" \" ",
"How far are we from I-88?\" \" ",
"Freaking perv.\" \" ",
"What?\" \"",
"At least five or six hours away.\" \"",
"You hungry, sweetie?\" \"",
"Why couldn't we have gone through Santa Fe?\" \"",
"Could you put a cork in it.\" \"",
"Well, if I do say so myself, ma'am, your husband here's quite right.\" \"",
"This is the only southbound road going to I-88.\" \"",
"From there, you can take the 40 to California but it's a long, old drive.\" \"",
"That doesn't bother me.\" \"",
"I like to drive at night.\" \"",
"I used to work the night patrol for 15 years.\" \" ",
"Is that right?\" \"",
"You a cop?\" \" ",
"Detective.\" \"",
"Not anymore, thank God.\" \"",
"I'm starting my own security firm.\" \"",
"You a cop too, mister?\" \"",
"No.\" \"",
"I'm in telecommunications.\" \"",
"I sell cell phones.\" \"",
"Well, hell, you won't find any customers around here.\" \"",
"Yeah, no kidding.\" \"",
"Everything's fine.\" \"",
"Oil's okay.\" \"",
"Beauty, come back here!\" \"",
"Hey!\" \"",
"Buddy, what do I owe you?\" \" ",
"Forty-six dollars, even.\" \" ",
"Bob, here, let me take care of that.\" \"",
"I can still afford the gas, Bukowski.\" \" ",
"Keep the change, my friend.\" \" ",
"That's not what I meant.\" \"",
"Beauty?\" \"",
"Beauty?\" \"",
"Beauty?\" \"",
"Beauty?\" \"",
"Come on, girl.\" \"",
"Beauty?\" \"",
"Beauty, come on, girl.\" \"",
"Hello?\" \"",
"Is anyone there?\" \"",
"Beauty.\" \" ",
"Beauty, what are you doing?\" \" ",
"Can I help you, miss?\" \"",
"I'm sorry.\" \"",
"It's just our dog.\" \"",
"She just\" \"She got out and she won't stay put.\" \" ",
"Lynn!\" \"",
"We're leaving!\" \" ",
"Come on, girl.\" \"",
"Let's go.\" \" ",
"That's all right.\" \"",
"I got her.\" \" ",
"Okay.\" \"",
"Honey?\" \"",
"Honey?\" \"",
"Could you tell him to take it a bit easy?\" \" ",
"The heat's making me nauseous.\" \" ",
"Let's go!\" \"",
"Let's go.\" \"",
"Let's go.\" \"",
"Let's go.\" \"",
"Honey, we're on vacation.\" \" ",
"Yeah?\" \" ",
"You know it's not on the map but if you take a left at the dirt road couple miles from here it'll lead you through the hills, probably save you a couple of hours.\" \"",
"Can't miss it.\" \"",
"There's an old fence right in front.\" \"",
"Well, thank you.\" \"",
"Thank you very much.\" \"",
"Old fence, dirt road, two miles.\" \" ",
"You have a safe trip.\" \" ",
"Thank you.\" \"",
"Dad, I hate your singing.\" \"",
"Guys, no.\" \"",
"Can you imagine the first people to cross this desert?\" \" ",
"They didn't know where they were.\" \" ",
"I can, because neither do we.\" \"",
"Brenda, the road is here, right?\" \"",
"It exists.\" \"",
"It's got to go somewhere.\" \"",
"Besides, look how beautiful it is.\" \"",
"The light on the rocks.\" \"",
"It's gorgeous.\" \"",
"This is so fucked.\" \"",
"Excuse me.\" \"",
"Watch your language, young lady.\" \"",
"I would like a little backup here.\" \"",
"This is such bullshit.\" \" ",
"Hey, watch your fucking mouth.\" \" ",
"Can we just calm down here?\" \" ",
"We're all a little tired, okay?\" \" ",
"Yeah, Mom.\" \"",
"Didn't you grow up in the '60s?\" \"",
"Why are you so uptight?\" \"",
"Your mom didn't used to be uptight.\" \"",
"She was a little hippie chick.\" \" ",
"No.\" \" ",
"Yeah.\" \"",
"You should've seen her at your age.\" \"",
"She was a little hottie.\" \" ",
"Dad.\" \" ",
"Stop that.\" \"",
"Driving through the New Mexico desert in the middle of summer with a broken air conditioner.\" \"",
"Great vacation.\" \"",
"Well, I'm glad you came because I would have been so fucking bored otherwise.\" \"",
"Yeah.\" \"",
"So bored.\" \"",
"Give up on that, man.\" \"",
"I tried for an hour and a half.\" \"",
"Thing's ancient.\" \"",
"Wiring's medieval.\" \"",
"Needs to be replaced.\" \"",
"There.\" \"",
"There.\" \"",
"It's just the thermostat.\" \" ",
"It was disconnected.\" \" ",
"Anyway at least we can breathe now.\" \"",
"Hey, I thought you quit smoking.\" \" ",
"Does my sister know?\" \" ",
"Hey, man fuck your sister.\" \"",
"You know, Big Bob would get pretty pissed off if he found out you were smoking in his newly renovated '88 Airstream.\" \"",
"Yeah?\" \"",
"Well, fuck Big Bob too.\" \"",
"Here.\" \"",
"Want one?\" \"",
"Go ahead.\" \"",
"Your sister would out my nuts off if you started.\" \"",
"I think she did that a long time ago.\" \"",
"I think you're right.\" \"",
"I think we're on this blue road.\" \"",
"Mom, we're not on the blue road or any other road.\" \"",
"The guy said it wasn't on the map, remember?\" \"",
"Oh, my God!\" \"",
"Hang on!\" \"",
"Watch the baby!\" \"",
"Honey?\" \"",
"Where's the baby?\" \"",
"Where's the baby?\" \"",
"She's right here.\" \"",
"She's fine.\" \" ",
"What the hell?\" \" ",
"What the hell happened?\" \"",
"The heat must have caused a blowout.\" \" ",
"Are you okay?\" \"",
"You sure?\" \" ",
"Yes, I'm fine.\" \"",
"I'm just a little shaken up.\" \" ",
"God... .\" \" ",
"Where's Bobby?\" \"",
"Bobby?\" \"",
"Are you all right, honey?\" \"",
"Are you okay?\" \"",
"He's all right.\" \"",
"I knew this wasn't safe for the baby.\" \"",
"We're okay!\" \"",
"Fine!\" \"",
"Son of a bitch!\" \"",
"Okay, okay.\" \"",
"We're all right.\" \" ",
"The whole trailer\" \" I know, I know.\" \"",
"Come here, guys.\" \"",
"Come here.\" \"",
"Beauty!\" \"",
"Beasfl\" \"We are so fucked.\" \" ",
"Did you find a signal?\" \" ",
"No.\" \"",
"Nothing.\" \"",
"Ninety-seven percent nationwide coverage.\" \" ",
"We find ourselves in that 3 percent.\" \" ",
"Yeah.\" \"",
"Your dad wanted to see the desert.\" \"",
"Hope he's enjoying himself.\" \"",
"Doug, don't start.\" \" ",
"Maybe somebody will need a rent-a-cop.\" \" ",
"Doug.\" \" ",
"What?\" \" ",
"Come on.\" \"",
"Every chance he gets, he takes me down a peg.\" \"",
"I can't snap back at him?\" \"",
"Yeah, well, don't stoop to his level.\" \"",
"Two hundred degrees out here.\" \"",
"It's everybody's fault.\" \"",
"It's all of you.\" \"",
"Get off my back.\" \"",
"General?\" \"",
"What's the damage?\" \"",
"The axle is broken.\" \"",
"And it's totaled.\" \"",
"Can we fix it?\" \"",
"What?\" \"",
"The frame is twisted to hell.\" \"",
"We can't\" \"With what?\" \"",
"I mean\" \"Give me your cell phone.\" \"",
"I'll call a tow truck.\" \"",
"No signal.\" \"",
"I tried.\" \"",
"Well, we can use the CB.\" \"",
"Try, but I doubt you're gonna get anybody.\" \"",
"The freeway's on the other side of those hills.\" \"",
"You won't get a signal.\" \"",
"So, what are we gonna do?\" \"",
"Excuse me.\" \"",
"Bob?\" \"",
"What are we doing?\" \"",
"We walk.\" \" ",
"We walk?\" \" ",
"Yeah.\" \"",
"You and me.\" \"",
"We're in the middle of the desert.\" \"",
"We're miles from anywhere.\" \"",
"Well, it's either that or we sit and wait for somebody to drive by.\" \"",
"Seen anybody drive by since we got off the freeway?\" \"",
"I haven't seen anyone since you took the shortcut.\" \" ",
"Honey, let's go for a walk.\" \" ",
"Okay.\" \"",
"There's nothing there, you shitheads.\" \"",
"I'm sure this desert is full of rattlesnakes.\" \" ",
"I think that's locked.\" \" ",
"Bobby, come here.\" \" ",
"Yes, sir.\" \" ",
"Thanks, honey.\" \"",
"You're welcome.\" \" ",
"You know, I saw\" \" Mom?\" \" ",
"What?\" \" ",
"Safety's on, boy.\" \" ",
"Yes, sir.\" \"",
"I saw a documentary on desert snakes on the Discovery Channel.\" \"",
"There's some species if you get bit, you die within, like, 25 minutes.\" \"",
"Really?\" \"",
"I'd like to see how they stack up against this one.\" \"",
"I thought this was gonna stay looked up, honey.\" \" ",
"I thought you weren't talking to me.\" \" ",
"You're right.\" \"",
"Relax, I'm a licensed professional.\" \"",
"I'll take my bullets over your prayers.\" \"",
"Some things never change.\" \"",
"So what about, like, scorpions and coyotes and stuff?\" \"",
"Praise the Lord, and pass the ammunition.\" \" ",
"Hey, Doug.\" \"",
"You wanna try?\" \" ",
"Hey!\" \"",
"Don't point that at me!\" \"",
"Don't worry.\" \"",
"The safety's on.\" \" ",
"Bobby?\" \" ",
"Yeah?\" \"",
"Leave Doug alone.\" \"",
"He's a Democrat.\" \"",
"He doesn't believe in guns.\" \"",
"Oh, come on, Dougie.\" \"",
"Come here.\" \"",
"Just one shot.\" \"",
"It makes you feel kind of powerful.\" \"",
"You might like it.\" \"",
"I don't think so.\" \"",
"I'd probably just shoot my foot off.\" \"",
"You know, he's right.\" \"",
"He'd probably shoot his foot off.\" \" ",
"Hey, B.\" \" Hey.\" \"",
"How you holding up?\" \"",
"I am thrilled.\" \"",
"Yeah.\" \"",
"This is a total drag.\" \"",
"Yeah, well, you know, I really don't care what they say.\" \"",
"Next year, I am going to CGHCUH with my friends.\" \"",
"Not going on any more of these family trips.\" \"",
"Well, we're not gonna have many more of them, you know?\" \"",
"And if you wanna go to CGHCUH, you're gonna have to get a job.\" \"",
"Oh, yeah.\" \"",
"What, you mean like your job?\" \"",
"Brenda, I help Doug out at the store, okay?\" \"",
"Yeah.\" \"",
"Honey, can you bring me my jacket?\" \"",
"I'll be right there, honey!\" \"",
"Bobby!\" \"",
"FuckeH\" \" Could you get me a Twinkie?\" \" ",
"No.\" \"",
"I'll walk in this direction.\" \"",
"Fine.\" \"",
"Find out where the road leads, I'll head back the way we came.\" \" ",
"Stay away from me.\" \" ",
"All right.\" \"",
"Road's gotta lead somewhere.\" \" ",
"Maybe to the freeway.\" \" ",
"Are you taking a gun too?\" \"",
"No.\" \"",
"No, honey, I'm not.\" \"",
"I don't need a gun to find help.\" \" ",
"No, I don't want this.\" \" ",
"You need the hat.\" \" ",
"You're walking in the desert.\" \" ",
"It gives me hat head.\" \" ",
"It's not about fashion.\" \" ",
"All right.\" \"",
"Thank you.\" \"",
"Listen, if you don't find help within five or six miles, just come back.\" \" ",
"Here, honey.\" \" ",
"I'm gonna go to the gas station.\" \"",
"You're just never gonna make it back by sunset is the only problem.\" \"",
"Honey, it's only about eight miles.\" \"",
"I'll borrow the old man's car, use his phone.\" \"",
"I'll be back with a tow truck before nightfall.\" \" ",
"Can I go with Doug?\" \" ",
"No.\" \"",
"Stay with your mother and sisters.\" \"",
"Now, Brenda, listen up.\" \"",
"Your brother is in charge, all right?\" \" ",
"So don't tease him.\" \" ",
"We should pray before you go.\" \" ",
"Oh, Mom!\" \" ",
"Brenda, is it really so much to ask?\" \" ",
"Yes.\" \" ",
"Come on.\" \" ",
"Damn it.\" \" ",
"We're waiting.\" \"",
"Thank God no one's watching us.\" \" ",
"Bobby.\" \"",
"Stop.\" \" ",
"Stop it.\" \"",
"Please bless us at this time, and send your guardian angels to help us to walk through this valley, in our faith with the help of our dear Lord.\" \"",
"Amen.\" \" ",
"Amen.\" \" ",
"Amen.\" \"",
"Okay.\" \" ",
"Bye, Dad.\" \" ",
"Be careful.\" \"",
"See you guys in a few hours.\" \"",
"Keep the hat on, okay?\" \" ",
"Am I wearing it?\" \" ",
"Don't give me that look.\" \" ",
"Do you see it on my head?\" \" ",
"I love you.\" \"",
"Hi.\" \"",
"Hi, sweetness.\" \"",
"Hello.\" \"",
"Who doesn't wanna sleep anymore?\" \"",
"Are you hungry?\" \"",
"Please take your feet off the table.\" \"",
"Honestly.\" \"",
"Just because we're camping doesn't mean you get to act like a pig.\" \"",
"Honey, really, you shouldn't go from the breast to bottle, breast to bottle.\" \"",
"At her age, she should get used to one or the other.\" \"",
"Mom, thanks for the advice, but she's my baby.\" \"",
"I can't believe we're stranded in the middle of nowhere on your anniversary.\" \"",
"If we just stayed on the main road, we'd be in California right now.\" \"",
"What I'd give for a hot shower and a cold margarita.\" \"",
"The beach.\" \" ",
"A massage.\" \" ",
"I'll take a real bed.\" \" ",
"Yeah.\" \" ",
"The chronic.\" \" ",
"What?\" \" ",
"Brenda.\" \" ",
"What?\" \" ",
"The chronic...\" \"It's pot, Mom.\" \" ",
"Who smokes pot?\" \" ",
"Yes, exactly, who smokes pot?\" \"",
"Hey, has anybody seen my red sweatshirt?\" \"",
"You know, the hoodie thing in the back seat?\" \"",
"Brenda?\" \"",
"Weren't you, like, sleeping on it?\" \"",
"I haven't touched your smelly sweatshirt.\" \"",
"Why are the dogs so excited?\" \"",
"There's probably some rattlesnakes somewhere around.\" \"",
"You know how Freud would have interpreted your obsession with rattlesnakes, Mom?\" \"",
"Bobby.\" \"",
"So bad.\" \" ",
"Bobby Carter!\" \"",
"Stop that.\" \" ",
"Gross.\" \"",
"So wrong.\" \"",
"That was a little gross.\" \"",
"I'm sorry.\" \"",
"I apologize to the table.\" \"",
"Well, thank you.\" \"",
"I'm really glad that we all came on this trip together.\" \"",
"Well, that makes one of us.\" \" ",
"Where is the fruit?\" \" ",
"Does anyone need anything?\" \"",
"No.\" \"",
"Brenda?\" \"",
"Are you kidding me?\" \"!\" \"",
"Do you know how long--?\" \" ",
"Bobby!\" \" ",
"Sorry.\" \" ",
"Bobby, it's dinnertime!\" \" ",
"I'll get him, Mom.\" \" ",
"This always happens.\" \" ",
"Bobby!\" \"",
"I don't want you going away from the trailer right now.\" \"",
"Beauty!\" \"",
"Beauty!\" \"",
"Beauty!\" \"",
"Beauty!\" \"",
"Fuck!\" \"",
"What the hell?\" \"",
"Look at that.\" \"",
"Hello?\" \"",
"Hello?\" \"",
"Hey,buddy?\" \"",
"Hello.\" \"",
"Hello!\" \"",
"Hey, I had a bit of a wreck out on your shortcut.\" \"",
"Yo!\" \"",
"I got some water, all right?\" \"",
"It's dead.\" \"",
"Damn.\" \"",
"Yo, my friend!\" \"",
"Hey!\" \"",
"You here?\" \"",
"You home?\" \"",
"Hello?\" \"",
"Hello?\" \"",
"You son of a bitch.\" \"",
"Yeah.\" \"",
"Jupiter.\" \"",
"I got a buckshot, you hear?\" \"",
"Hey, mister.\" \"",
"You don't understand what's going on here.\" \"",
"Hey!\" \"",
"The kids grew up in the mines like wild animals.\" \"",
"Open the door, slowly!\" \"",
"What kind of place is that for children?\" \"",
"What kind of a place is that?\" \"",
"Okay.\" \"",
"Put the weapon down.\" \"",
"Put it down, easy.\" \"",
"Put it down.\" \"",
"I did the best I could.\" \" ",
"I'm sorry.\" \" ",
"Hey!\" \"",
"Shit.\" \"",
"Daddy.\" \"",
"Daddy.\" \"",
"Daddy.\" \"",
"Daddy.\" \"",
"Daddy.\" \"",
"Daddy.\" \"",
"Daddy.\" \"",
"Daddy.\" \"",
"Would you give me that gun, Pluto!\" \" ",
"Give me the darn gun.\" \" ",
"Get over here.\" \" ",
"That's mine.\" \" ",
"Come on!\" \"",
"You oughtn't to have a gun.\" \"",
"Brenda!\" \"",
"Brenda!\" \"",
"Hey, I'm here!\" \"",
"I'll be back in a minute!\" \"",
"Bobby!\" \"",
"Bobby?\" \"",
"Bobby.\" \"",
"Bobby.\" \"",
"Bobby!\" \"",
"Okay, let me just-- Oh, honey, the hair.\" \"",
"Just...\" \"You know, you really are so lucky that Brenda found you.\" \"",
"I mean, what if you'd had to spend the whole night out there in the cold?\" \"",
"Hey, Bobby.\" \"",
"Don't worry.\" \"",
"Beauty will come back when she's hungry, okay?\" \" ",
"Did you try the radio?\" \" ",
"It doesn't work.\" \"",
"But we heard something.\" \"",
"Some sort of breathing.\" \"",
"What breathing?\" \"",
"It was weird.\" \"",
"Right, Mom?\" \"",
"I'm sure it was just static.\" \"",
"Mom!\" \"",
"It sounded like a perverted call.\" \"",
"Only Mom would get an obscene phone call out here.\" \"",
"Bobby.\" \"",
"What--?\" \" ",
"Honey?\" \" ",
"No, no!\" \"",
"I want you guys to stay here.\" \"",
"What is wrong with him?\" \"",
"What's he doing?\" \"",
"Shit.\" \"",
"Beasfl\" \" Beasfl\" \" Hey,Bobby\" \" Jesus Christ!\" \" ",
"Hey, whoa!\" \"",
"Stop pointing the gun at me.\" \"",
"Please.\" \" ",
"Jesus, Bobby.\" \" ",
"Doug, I'm sorry, man.\" \"",
"It's me.\" \" ",
"You okay?\" \" ",
"Yeah.\" \"",
"God.\" \"",
"I didn't mean to startle you, man.\" \"",
"I found this place.\" \"",
"You wouldn't believe it.\" \"",
"It's like The Twilight Zone.\" \" ",
"Hey.\" \" ",
"Is that Bob?\" \" ",
"It's Doug.\" \"",
"It's Doug.\" \" ",
"It's okay.\" \"",
"Oh, honey.\" \"",
"Oh, my God.\" \"",
"Thank God you're okay.\" \"",
"I was so worried.\" \"",
"No, I'm fine.\" \"",
"Just exhausted.\" \" ",
"Look at all this stuff I found.\" \" ",
"What is this?\" \"",
"There's a huge crater out there.\" \"",
"People must use it as a dumpsite.\" \"",
"I've never seen anything like it in my life.\" \"",
"A lot of this stuff is brand-new.\" \"",
"What are we gonna do with a fishing pole out here?\" \" ",
"I thought you went to find somebody.\" \" ",
"The road dead-ends in five miles.\" \"",
"That's where the crater is.\" \"",
"It's not a shortcut.\" \" ",
"It dead-ends?\" \" ",
"Dead end.\" \" ",
"That old man was full of shit.\" \" ",
"I hope your dad did better.\" \"",
"Maybe we just missed a turn.\" \"",
"You guys cook any dinner?\" \"",
"I'm starving.\" \" ",
"Yeah.\" \"",
"Come on in.\" \" ",
"Great.\" \"",
"Thanks.\" \"",
"Brenda, you have any lip balm?\" \"",
"My lips are really dry.\" \"",
"Are you serious?\" \"",
"That's gross.\" \"",
"I'm not letting you use my lip balm.\" \"",
"Do you use it without telling me?\" \" ",
"There's nothing wrong with him\" \" Baby needs to go to bed.\" \"",
"Wait, Bobby, don't close that door.\" \"",
"We're leaving in a second.\" \"",
"Get some rest with Daddy.\" \"",
"You guys aren't gonna wait till Dad gets back?\" \"",
"No, Bobby, I'm exhausted.\" \"",
"I need to get some sleep.\" \"",
"No, no, no.\" \"",
"It's fine.\" \"",
"You'll be more comfortable here.\" \"",
"Because I'll sleep on the floor.\" \"",
"Mom and Brenda will be up front, you and Lynn in back.\" \"",
"We got extra sleeping bags and stuff too.\" \"",
"Bobby, you got six people sleeping in a three-person trailer.\" \"",
"It's not comfortable.\" \"",
"It's a little bit weird.\" \"",
"Okay?\" \" ",
"Honey, you gonna bring the water?\" \" ",
"Yeah.\" \" ",
"Hey, Doug.\" \"",
"Can I talk to you?\" \" ",
"What's up?\" \" ",
"Baby, you coming?\" \" ",
"I'm almost ready.\" \"",
"What's going on?\" \"",
"When I...\" \" Bobby.",
"Bobby.\" \" ",
"No,no,no.\" \"",
"Hey, I'm sure your dad's gonna be back soon, okay?\" \"",
"I mean, come on.\" \"",
"He's Big Bob, right?\" \"",
"It's gonna take more than a couple of snakes and scorpions to stop him.\" \"",
"Okay?\" \" ",
"Excuse me.\" \" ",
"Sorry.\" \"",
"Did you get pillows?\" \"",
"They're in the SUV.\" \"",
"Hey, hey, listen.\" \"",
"If he's not back by midnight, we'll both go looking for him.\" \"",
"Deal?\" \" ",
"Yeah.\" \"",
"Deal.\" \" ",
"Okay.\" \" ",
"Get some rest, man.\" \" ",
"Good night, Bobby.\" \"",
"Good night.\" \"",
"What are you doing?\" \"",
"There's no mosquitoes in the desert.\" \"",
"Honey, there's mosquitoes everywhere.\" \"",
"Hi.\" \"",
"Beast\" \"Beast?\" \"",
"Beasfl\" \"Beast\" \"Beast\" \"Beast\" \"Beasty?\" \"",
"Come here, boy.\" \"",
"Beast?\" \"",
"GUYS!\" \"",
"Guys!\" \" ",
"Oh, God.\" \" ",
"Guys.\" \"",
"Open.\" \" ",
"Jesus.\" \" ",
"Bobby\" \"What is it, Bobby?\" \" ",
"Something is going on around here.\" \" ",
"What is it?\" \"",
"There are people or something living in those hills.\" \" ",
"People?\" \" ",
"No, no, no.\" \"",
"Doug.\" \" ",
"Doug, please.\" \" ",
"Bobby.\" \"",
"Bobby, listen to me.\" \"",
"We're in the middle of nowhere, okay?\" \"",
"Nobody lives in those hills.\" \"",
"Beauty's dead.\" \" ",
"What?\" \" ",
"I found her and she was, like...\" \"It looked like somebody had, like, out her open, you know?\" \" ",
"Cut her open?\" \" ",
"Yeah.\" \"",
"I didn't want to say anything because I didn't want to scare everyone and I didn't wanna get Mom upset.\" \"",
"But I'm telling you, there is something going on, okay?\" \"",
"And Dad's not even back yet.\" \"",
"Okay.\" \"",
"All right.\" \"",
"Stay right there.\" \"",
"I'll come check this out.\" \"",
"We're not alone.\" \"",
"All right, Lynn, you stay here, you lock the car, okay?\" \"",
"We're gonna check this out.\" \"",
"Now, where did you see this?\" \"",
"Now!\" \" ",
"It's Bob.\" \" ",
"Wait.\" \"",
"We don't know what that is.\" \" ",
"It's Bob!\" \" ",
"We don't know what that is!\" \" ",
"Bob!\" \"",
"I'm coming.\" \" ",
"Dad?\" \"",
"Dad!\" \"",
"Help!\" \"",
"Brenda, watch the baby.\" \"",
"No!\" \"",
"Get back!\" \"",
"Hey, little birdie.\" \"",
"Hey, little birdie.\" \"",
"Get your mother out of here!\" \"",
"Get your mother out of here!\" \"",
"Doug!\" \"",
"Do something!\" \"",
"You've gotta be a man to do that!\" \"",
"I told you to move, so move!\" \"",
"Get!\" \"",
"Get!\" \"",
"Fuck off!\" \"",
"Not my Bob!\" \" ",
"It's not my Bob!\" \" ",
"Come on, Mom.\" \"",
"Let's go.\" \"",
"No, just come on.\" \"",
"What was that?\" \"",
"Mom, just stay here, okay?\" \"",
"Help!\" \"",
"Help!\" \"",
"Oh, you fat.\" \"",
"Big, fat and juicy.\" \"",
"No.\" \"",
"No, please!\" \"",
"Bobby!\" \"",
"Bobby, where you going?\" \"",
"Fucking kill those motherfuckers!\" \"",
"Get back here!\" \"",
"Don't do something stupid!\" \"",
"They're in the trailer!\" \" ",
"Get her out of the door!\" \" ",
"Help!\" \"",
"Stop it!\" \"",
"Shoot him!\" \"",
"Brenda!\" \" ",
"Help me!\" \" ",
"Come on, now.\" \" ",
"Look at me.\" \"",
"Look at me.\" \" ",
"Stop, stop.\" \"",
"That's it.\" \"",
"Look at me.\" \"",
"Shit!\" \" ",
"I'll come back for you.\" \" ",
"Brenda!\" \"",
"Bitch stuck me like a pig!\" \"",
"Are you okay?\" \"",
"Are you okay?\" \"",
"Oh, God!\" \"",
"Don't move.\" \"",
"Don't move, honey.\" \"",
"Don't move.\" \"",
"Okay.\" \"",
"Okay.\" \"",
"All right.\" \"",
"You're gonna be okay.\" \"",
"You're gonna be okay.\" \"",
"Sweetheart.\" \"",
"Okay.\" \"",
"Honey?\" \"",
"Sweetheart?\" \"",
"Lynn, come on!\" \"",
"Lynn!\" \"",
"Come on.\" \"",
"What are we gonna do, huh?\" \"",
"What are we gonna do if they come back, Doug?\" \"",
"This trailer is so small.\" \"",
"I clean and clean.\" \"",
"It's still a mess.\" \"",
"It's a mess.\" \"",
"Is Big Bob home?\" \"",
"Not yet.\" \"",
"Everybody went to bed?\" \" ",
"Yes?\" \" ",
"Yeah.\" \"",
"Brenda's right here and Bobby's in the back.\" \"",
"Lynnie's sleeping?\" \"",
"You need anything?\" \"",
"A blanket.\" \" ",
"It's cold.\" \"",
"It's cold.\" \" ",
"Okay.\" \"",
"Nice and warm, huh?\" \"",
"Nice and warm.\" \"",
"You're so sweet, Doug.\" \"",
"I understand why Lynnie loves you so much.\" \"",
"I'm not gonna take this shit.\" \"",
"We're not going anywhere.\" \"",
"You wanna die too?\" \" ",
"I don't fucking care!\" \" ",
"Do you wanna die?\" \"!\" \"",
"Look what they did to your mother!\" \"",
"Look what they did to your sister!\" \"",
"You need to think.\" \"",
"You're not a child anymore.\" \"",
"You won't make it three feet out there.\" \"",
"Now, we need a plan.\" \"",
"We need to think.\" \" ",
"We need to think.\" \" ",
"Think about what?\" \"!\" \"",
"They have Catherine, your daughter, and you're gonna sit and do nothing?\" \"!\" \"",
"You're a pussy, just like my dad said!\" \"",
"You shut the fuck up!\" \"",
"Shut up!\" \"",
"Shut your mouth!\" \"",
"Fuck you.\" \"",
"Goggle?\" \"'",
"G°9Q/e?\" \" ",
"No!\" \"",
"Turn the lights on.\" \"",
"Turn the lights on.\" \"",
"Turn the lights on.\" \" ",
"He said he's coming back.\" \" ",
"Quiet.\" \"",
"He said he's coming back for me.\" \"",
"Please.\" \"",
"It's them.\" \"",
"It's them.\" \"",
"It's them.\" \"",
"Goggle?\" \"",
"Goggle?\" \"",
"You copy, Goggle?\" \"",
"Where's Goggle?\" \"",
"He stayed back there.\" \"",
"He's keeping an eye on them.\" \"",
"I told you.\" \"",
"What?\" \"",
"I told you, you've gotta kill them all!\" \"",
"Beast!\" \"",
"Beast!\" \"",
"You're alive!\" \"",
"Oh, good boy.\" \"",
"Good Beast.\" \"",
"Oh, good boy.\" \"",
"Good boy.\" \"",
"Jupiter's going to finish them off.\" \"",
"Why are you doing this to us?\" \"",
"Why are you doing this?\" \"",
"You hear me?\" \"",
"Give me back my baby.\" \"",
"What the fuck do you want from us?\" \"",
"What do you want?\" \"!\" \"",
"You give me back my little girl, you hear me?\" \"",
"Bobby how many bullets do you have left?\" \"",
"A clip and a half.\" \"",
"What are we doing?\" \"",
"Which way, Beast?\" \"",
"Which way?\" \"",
"Come on.\" \"",
"Follow the trail.\" \"",
"Come on, big guy\" \"I hope you're not afraid of the dark.\" \"",
"Where are we?\" \"",
"Which way, Beast?\" \"",
"Good boy.\" \"",
"Bobby?\" \"",
"Bobby, I followed the blood trail.\" \"",
"It leads to a town or something out here.\" \"",
"Bobby?\" \"",
"Who knows, maybe someone heard our calls.\" \"",
"Nobody's gonna hear the fucking calls, Brenda!\" \"",
"Nobody will see the goddamn smoke in this desert!\" \"",
"We're in the middle of freaking\" \"Shit!\" \" ",
"I'm sorry.\" \" ",
"It's okay.\" \"",
"I'm so scared.\" \"",
"Who are these people?\" \"",
"Good dog.\" \"",
"Stay.\" \" ",
"Did you push her?\" \" ",
"Yes.\" \" ",
"Did you destroy the furniture?\" \" ",
"No.\" \" ",
"Did you call her trailer-park trash?\" \" ",
"I did not.\" \"",
"I did treasure my marriage.\" \"",
"I respected her.\" \"",
"That's when her abuse started.\" \" ",
"So now you're saying\" \" I'm still in love with her.\" \"",
"You couldn't \"treasure the marriage\" if you wanted to end it two weeks into it.\" \"",
"I wanted to end it because I had a feeling that things weren't gonna work.\" \"",
"She wasn't working as a team player.\" \" ",
"I tried coaching her\" \" What were you doing coaching her?\" \" ",
"Is she your child?\" \"!\" \" ",
"No, she's not my child\" \"Coaching her in what?\" \"",
"She wanted me to eat dinner, spaghetti and I wanted to go out before it got dark.\" \"",
"It was getting dark\" \" She hit me with a plate of spaghetti.\" \" ",
"She hit you?\" \"",
"When that happened, I grabbed my coat and I ran out the door.\" \"",
"Watch her head, Bobby.\" \"",
"Come on.\" \"",
"O say, can you see\" \"By the dawn's early light\" \"What so proudly we hailed\" \"At the twilight's last gleaming?\" \"",
"Whose broad stripes and bright stars\" \"Through the perilous fight\" \"O'er the ramparts we hailed\" \"Were so gallantly streaming?\" \"",
"And the rockets' red glare\" \"The bombs bursting in air\" \"Gave proof through the night\" \"That our flag was still there\" \"Where's my daughter?\" \"",
"Where is she?\" \"",
"Answer me.\" \"",
"I don't know where she is.\" \"",
"I never leave this place.\" \"",
"Your people asked our families to leave their town.\" \"",
"And you destroyed our homes.\" \"",
"We went into the mines.\" \"",
"You set off your bombs and turned everything to ashes.\" \"",
"You made us what we've become.\" \"",
"Boom.\" \"",
"Boom.\" \"",
"Boom.\" \"",
"Where's my baby?\" \"",
"What's so funny?\" \"",
"It's breakfast time.\" \" ",
"Please don't kill me.\" \"\"",
"Please don't kill me.\"\" \" ",
"Please don't kill me.\" \"\"",
"Please don't kill me.\"\" \"",
"Lizard.\" \"",
"Kill the baby.\" \"",
"Baby.\" \"",
"Baby.\" \"",
"Get out the way, Ruby.\" \"",
"I said, get.\" \"",
"Now get.\" \"",
"Mister, would you play with us?\" \"",
"Ruby!\" \"",
"Catherine!\" \" ",
"It's okay.\" \" ",
"Fine.\" \"",
"It's okay.\" \"",
"It's okay.\" \"",
"It's okay.\" \"",
"No.\" \"",
"No.\" \"",
"No.\" \" ",
"No!\" \" ",
"Come on, Brenda.\" \" ",
"Bobby!\" \" ",
"Stay with me.\" \"",
"Stay with me.\" \" ",
"What happened?\" \" ",
"Come on.\" \"",
"Come on.\" \"",
"It's gonna be all right.\" \"",
"Careful.\" \"",
"Please.\" \"",
"Please.\" \"",
"Shit.\" \"",
"Come on out, you motherfucker.\" \"",
"Look.\" \"",
"Look.\" \"",
"Look.\" \"",
"It's just a tumbleweed.\" \"",
"Come on.\" \"",
"Come on, let's go back to the trailer.\" \"",
"Come on.\" \"",
"Come on.\" \" ",
"Oh, my God!\" \" ",
"She's gone.\" \"",
"She's fucking gone!\" \"",
"She's gone!\" \"",
"Brenda!\" \" ",
"Go get the trailer ready!\" \"",
"Brenda!\" \"",
"Go\" \" Where's Mom?\" \"!\" \" ",
"Go get the trailer ready, okay?\" \"",
"Go.\" \" ",
"Where's Mom?\" \"!\" \"",
"Where's Mom?\" \"!\" \"",
"Listen, go get the trailer ready.\" \"",
"Go in.\" \"",
"Do what we talked about.\" \"",
"Go, go, go.\" \"",
"Get the window.\" \"",
"Get the window.\" \"",
"It's stuck!\" \"",
"Move!\" \"",
"Go, go, go.\" \"",
"Get out.\" \"",
"Here.\" \"",
"Ruby!\" \"",
"Ruby!\" \"",
"Ruby!\" \"",
"Ruby!\" \"",
"Come back here!\" \"",
"You little bitch!\" \"",
"Come back!\" \"",
"Give me that baby!\" \"",
"Fucker!\" \"",
"Come on, let's get out of here.\" \"",
"Oh, my God.\" \"",
"Look!\" \"",
"Bobby!\" \"[",
"ENGLISH]\""
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[
"using HotChocolate.",
"Types;\nusing Snowflake.",
"Filesystem;\nusing System;\nusing System.",
"Collections.",
"Generic;\nusing System.",
"IO;\nusing System.",
"Linq;\nusing System.",
"Text;\n\nnamespace Snowflake.",
"Remoting.",
"GraphQL.Model.",
"Filesystem.",
"Contextual\n{\n public sealed class OSDirectoryContentsInterface\n : InterfaceType\n {\n protected override void Configure(IInterfaceTypeDescriptor descriptor)\n {\n descriptor.",
"Name(\"OSDirectoryContents\")\n .Description(\"Describes the contents of a directory in the realized, OS-dependent file system.\")",
"\n .Interface<DirectoryContentsInterface>();\n descriptor.",
"Field(\"root\")\n .Description(\"The root directory.\")",
"\n .Type<OSDirectoryInfoInterface>();\n descriptor.",
"Field(\"files\")\n .Description(\"The files contained in this directory.\")",
"\n .Type<NonNullType<ListType<NonNullType<OSFileInfoType>>>>();\n descriptor.",
"Field(\"directories\")\n .Description(\"The child directories contained in this directory.\")",
"\n .Type<NonNullType<ListType<NonNullType<OSDirectoryInfoInterface>>>>();\n descriptor.",
"Field(\"directoryCount\")\n .Description(\"The number of child directories contained in this directory.\")",
"\n .Type<NonNullType<IntType>>();\n descriptor.",
"Field(\"fileCount\")\n .Description(\"The number of child files contained in this directory.\")",
"\n .Type<NonNullType<IntType>>();\n }\n }\n}\n"
] | {
"pile_set_name": "Github"
} | [
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[
"Background {#Sec1}\n==========\n\nPain due to cancer is notoriously difficult to treat, adds substantially to the emotional burden of having cancer, and is more frequently observed with certain cancer types such as with head and neck carcinomas (HNC). ",
"Indeed, 70--85 % of HNC patients report pain as their major symptom \\[[@CR1]--[@CR4]\\]. ",
"Patients typically report spontaneous pain as well as movement-related pain (i.e., mechanical allodynia) at the primary site of tumor development such as the tongue, floor of the mouth, larynx, oropharynx, hypopharynx and salivary glands \\[[@CR5], [@CR6]\\]. ",
"Many patients subsequently require opiates and tolerance rapidly develops for many HNC pain patients \\[[@CR7]\\].",
"\n\nDespite the occurrence of pain in cancer, there is incomplete understanding of the mechanisms by which oral tumors evoke pain. ",
"Because pain is often the first symptom of oral cancer, occurring even when the tumor is still quite small in size, it is likely that oral cancer cells control the activities of surrounding nociceptors at the site of the tumor. ",
"A major class of trigeminal nociceptors express transient receptor potential (TRP) ion channels \\[[@CR8]--[@CR10]\\], including TRP vanilloid 1 (TRPV1), which has been implicated in oral cancer pain \\[[@CR7], [@CR11]\\]. ",
"However, the mechanism by which TRPV1 is activated during oral cancer is unknown.",
"\n\nMany polyunsaturated fatty acids (PUFA) are known to activate or sensitize TRPV1 \\[[@CR12]--[@CR19]\\]. ",
"However, no prior studies have evaluated the role of these lipids in mediating cancer pain. ",
"Interestingly, clinical studies have demonstrated that several TRP-active PUFAs such as linoleic acid, arachidonic acid, hydroxyoctadecadienoic acid (HODE) and hydroxyeicosatetraenoic acid (HETE) are significantly elevated in the saliva of oral cancer patients compared to normal patients, supporting a possible linkage between these lipids and oral cancer pain \\[[@CR20]\\]. ",
"In addition, TRPA1 is co-expressed with TRPV1 \\[[@CR21]\\], is also activated by oxidized lipids \\[[@CR12], [@CR22]--[@CR24]\\], modulates the activity of TRPV1 \\[[@CR25], [@CR26]\\] and may participate in cancer pain \\[[@CR27]\\]. ",
"Accordingly, we tested the novel hypothesis that oral squamous cell carcinomas release certain oxidized lipids that activate TRPV1 and TRPA1 on sensory neurons, contributing to the development of oral cancer pain.",
"\n\nMethods {#Sec2}\n=======\n\nCells lines {#Sec3}\n-----------\n\nHuman oral squamous cell carcinoma (OSCC) cell lines; HSC2, HSC3 and HSC4 were purchased from Health Science Research Resources Bank, Japan. ",
"These cell lines were established from tumors obtained from male patients with the primary site of tumor being either the floor of the mouth or the tongue \\[[@CR28]\\]. ",
"The OSCC cell lines were cultivated and maintained in Dulbecco's minimal essential medium (DMEM, Life Technologies, Carlsbad, CA) supplemented with glutamine, penicillin/streptomycin (pen-strep) and 10 % fetal bovine serum. ",
"The immortalized human normal oral keratinocyte (iNOK) cell line; OKF6-TERT2, was kindly provided by Dr. Cara Gonzales of UTHSCSA. ",
"This cell line was maintained in keratinocyte serum-free media (Life Technologies, Carlsbad, CA) supplemented with 25 ug/ml bovine pituitary extract (BPE), 0.2 ng/ml epidermal growth factor (EGF) and 0.4 uM calcium chloride.",
"\n\nAnimals {#Sec4}\n-------\n\nAll protocols were approved by the Institutional Animal Care and Use Committee of the University of Texas Health Science Center at San Antonio. ",
"Male Sprague--Dawley rats (Charles River Laboratories, Inc., Wilmington, MA, USA) were used for all experiments. ",
"Animals were housed for at least 7 days prior to the experiments.",
"\n\nDrugs {#Sec5}\n-----\n\nThe TRPV1 antagonist, AMG517 was purchased from Selleckchem (Houston, Texas, USA) and the TRPA1 antagonist, HC030031 was purchased from Tocris (Ellsville, Missouri, USA). ",
"Both drugs were diluted in 10 % DMSO/80 % mineral oil for peripheral injections and for systemic injections, AMG517 was diluted in 10 % DMSO/4 % Tween/PBS and HC030031 was diluted in 100 % DMSO.",
"\n\nPreparation of lipid extracts {#Sec6}\n-----------------------------\n\nCells (3 × 10^5^) were seeded per well of a 6-well plate. ",
"Twenty-four hours later, media was changed to 2 ml of basal minimal essential medium (MEM) without pH indicator, without FBS and supplemented with glutamine and pen-strep for OSCC cells and basal keratinocyte-serum free media with pen-strep but without BPE, EGF or calcium chloride. ",
"Following 24 hours, conditioned media was harvested and cells were trypsinized and counted using the TC20 automated cell counter (Bio-Rad). ",
"Conditioned media was subjected to lipid extraction using the Folch's Method \\[[@CR29]\\]. ",
"Briefly, 10 ml of chloroform: methanol (2:1) was added to 2 ml of conditioned media and vortexed for 10 sec and incubated on ice for 30 min. ",
"The solution was then vortexed again for 10 sec, washed with saline and centrifuged at 8000 RPM for 10 min. ",
"The lower chloroform layer was collected and dried under nitrogen gas and stored at −80 °C until further use. ",
"Light exposure was minimized during lipid extraction. ",
"Immediately prior to experiments, the dried extracts were re-suspended in phosphate buffered saline.",
"\n\nBehavioral testing {#Sec7}\n------------------\n\nAll observers were blinded to treatment allocation. ",
"Three behavioral assays were used in the study to measure spontaneous nocifensive as well as evoked nociception in response to noxious stimuli. ",
"Spontaneous nocifensive behavior was carried out as described \\[[@CR30]\\], whereas thermal allodynia was measured using the radiant heat test \\[[@CR31]\\] and mechanical allodynia was measured using the dynamic aesthesiometer as described previously \\[[@CR32]\\]. ",
"The effect of OSCC-released lipids on nociception was studied by injecting 50 ul of re-suspended lipid extract into the plantar surface of the rat hindpaw. ",
"Nocifensive behavior (defined as spontaneous flinching, flexion movements or licking of the treated hindpaw) was recorded every 2 min up to a total of 10 min. ",
"Thermal and mechanical thresholds were measured from 10 to 150 min after injection.",
"\n\nTo determine the role of TRPV1 and TRPA1 in mediating extract-induced nociceptive behaviors, animals were pretreated with vehicle, AMG517 or HC030031 45 min prior to extract injection. ",
"A systemic dose of 100 mg/kg of either drug, subcutaneously underneath the neck, was given for spontaneous nocifensive assays, whereas 100 ug of either drug was injected intraplantarly in the hindpaw for thermal and mechanical threshold measurements. ",
"The doses for each drug were selected based on previously published literature \\[[@CR33]--[@CR36]\\]. ",
"Nocifensive behavior was measured for 10 min whereas thermal and mechanical thresholds were measured at the peak time of allodynia; which was 10 min post injection of the lipid extract. ",
"Paw withdrawal thresholds for ipsilateral as well as contralateral hindpaws were measured.",
"\n\nStatistics {#Sec8}\n----------\n\nData are presented as mean ± SEM. ",
"Statistical analyses were performed using one-way or two-way ANOVA with Sidak's post-hoc test. ",
"A statistically significant difference was defined as *P* \\< 0.05. ",
"Error bars are standard error of the mean (S.E.M).",
"\n\nResults {#Sec9}\n=======\n\nOSCC-released lipids induce nocifensive behavior in rats in a TRPV1-dependent manner {#Sec10}\n------------------------------------------------------------------------------------\n\nBecause oral cancer patients report spontaneous pain \\[[@CR5], [@CR6]\\], we evaluated the role of OSCC-released lipids in triggering spontaneous nocifensive behavior in rats. ",
"Intraplantar injection of lipid extracts from OSCC cell lines HSC2, HSC3 and HSC4 evoked spontaneous nocifensive behaviors in a cell line-dependent manner (Fig.",
" [1b](#Fig1){ref-type=\"fig\"}). ",
"HSC2 produced the greatest response, followed by HSC3, whereas HSC4 failed to evoke a nocifensive response. ",
"The negative controls consisted of phosphate buffered saline (PBS; used to solubilize the lipid extracts) and lipid extracts from cell culture media without exposure to cells; neither of these controls evoked nocifensive behavior. ",
"To determine whether the effect of these lipids was specific to cancer cells and not simply a product of oral keratinocytes, we evaluated whether iNOK cells release lipids capable of producing nocifensive behavior. ",
"As seen in Fig.",
" [1a](#Fig1){ref-type=\"fig\"}, lipids extracts made from iNOK cells did not evoke a nocifensive response more than the extracts from the iNOK growth media alone. ",
"The iNOK growth media itself showed a nocifensive response that could be attributed to the basal media constituents required to maintain the cells. ",
"Collectively, these data indicate that the nocifensive effects of lipids are specific to OSCC cells.",
"Fig. ",
"1Effect of OSCC-released lipids on nocifensive behavior of rats. ",
"Lipid extracts using the Folch's extraction method were made from conditioned media of (**a**) immortalized normal oral keratinocyte (iNOK) cells and (**b**)*.* ",
"HSC2, HSC3 *and* HSC4 cells. ",
"50ul of re-suspended extracts were injected into the plantar surface of rat hindpaw and nocifensive behavior was measured upto 10 min post injection. ",
"N = 6/group. ",
"Vehicle group was injected with 50ul of PBS. ",
"Media group was injected with lipids extracted from media without exposure to cells. ",
"Data are represented as mean for each group and error bars indicate SEM. ",
"Data were analyzed using two-way ANOVA with Sidak's post-hoc test with p \\<0.05. ",
"Average cell numbers per well at the time of harvest of conditioned media are indicated. **",
"c** AMG517 (3 mg/kg) or HC030031 (30 mg/kg) was injected subcutaneously underneath the neck and 1 hour later, lipid extract from HSC2 cells was injected in the rat hindpaw and nocifensive behavior measured for 10 min. ",
"N =6/group. ",
"Data are represented as mean for each group and error bars indicate SEM. ",
"Data were analyzed using two-way ANOVA with Sidak's post-hoc test with p \\<0.05. ",
"Asterisks indicate \\**p* \\< 0.05; \\*\\**p* \\< 0.01; \\*\\*\\**p* \\< 0.001 and \\*\\*\\*\\**p* \\< 0.0001. ",
"Average cell numbers per well at the time of harvest of conditioned media are indicated\n\nTo determine whether OSCC-released lipids induce nocifensive behavior via activation of TRPV1 and/or TRPA1, rats were pretreated with a systemic dose of specific TRPV1 (AMG517, 3 mg/kg) or TRPA1 antagonist (HC030031, 30 mg/kg) prior to injection of HSC2 lipid extract. ",
"As seen in Fig.",
" [1c](#Fig1){ref-type=\"fig\"}, AMG517 pretreatment virtually abolished lipid-induced nocifensive behavior, whereas HC030031 pretreatment had no effect.",
"\n\nOSCC-released lipids induce thermal and mechanical allodynia in rats {#Sec11}\n--------------------------------------------------------------------\n\nWe next evaluated the effect of OSCC-derived lipids on thermal and mechanical thresholds. ",
"Figure [2](#Fig2){ref-type=\"fig\"}b, c and d demonstrated that injection of lipid extracts from HSC2 and HSC4 evoked significant thermal allodynia that lasted up to 50--90 min depending on the cell line. ",
"Lipid extracts from HSC3 did not produce a significant reduction in thermal escape thresholds. ",
"However, lipid extracts from all three-cell lines evoked significant mechanical allodynia that lasted up to 50--150 min in a cell line-dependent fashion (Fig.",
" [3b, c and d](#Fig3){ref-type=\"fig\"}). ",
"In contrast, lipids extracted from the iNOK cell line did not evoke significant thermal or mechanical allodynia compared to extracts made from growth media alone (Figs.",
" [2a](#Fig2){ref-type=\"fig\"} and [3a](#Fig3){ref-type=\"fig\"}).Fig. ",
"2Effect of OSCC-released lipids on thermal thresholds of rats. ",
"Lipid extracts using the Folch's extraction method were made from conditioned media of (**a**)*.* ",
"iNOK, (**b**)*.* ",
"HSC2, (**c**)*.* ",
"HSC3 and (**d**)*.* ",
"HSC4 cells. ",
"50ul of re-suspended extracts were injected into the plantar surface of rat hindpaw and thermal thresholds using the radiant heat test were measured up to 150 min post injection. ",
"N = 6/group. ",
"Vehicle group was injected with 50ul of PBS. ",
"Data are represented as mean for each group and error bars indicate SEM. ",
"Data were analyzed using two-way ANOVA with Sidak's post-hoc test with p \\<0.05. ",
"Asterisks indicate \\**p* \\< 0.05; \\*\\**p* \\< 0.01; \\*\\*\\**p* \\< 0.001 and \\*\\*\\*\\**p* \\< 0.0001. ",
"Average cell numbers per well at the time of harvest of conditioned media are indicatedFig. ",
"3Effect of OSCC-released lipids on mechanical thresholds of rats. ",
"Lipid extracts using the Folch's extraction method were made from conditioned media of (**a**)*.* ",
"iNOK, (**b**)*.* ",
"HSC2, (**c**)*.* ",
"HSC3 and (**d**)*.* ",
"HSC4 cells. ",
"50uls of re-suspended extracts were injected into the plantar surface of rat hindpaw and mechanical thresholds using the dynamic aesthesiometer were measured up to 170 min post injection. ",
"N =6/group. ",
"Vehicle group was injected with 50ul of PBS. ",
"Data are represented as mean for each group and error bars indicate SEM. ",
"Data were analyzed using two-way ANOVA with Sidak's post-hoc test with p \\<0.05. ",
"Asterisks indicate \\**p* \\< 0.05; \\*\\**p* \\< 0.01; \\*\\*\\**p* \\< 0.001 and \\*\\*\\*\\**p* \\< 0.0001. ",
"Average cell numbers per well at the time of harvest of conditioned media are indicated\n\nOSCC-released lipids mediate thermal but not mechanical allodynia via TRPV1 and TRPA1 channels {#Sec12}\n----------------------------------------------------------------------------------------------\n\nTo evaluate whether OSCC-released lipids regulate peripheral activities of TRP channels, rats were pre-treated with intraplantar injection of a vehicle, a TRPV1 antagonist (AMG517) or a TRPA1 antagonist (HC030031) followed by extract injection. ",
"Administration of the TRPV1 antagonist reversed lipid-evoked thermal allodynia by 93 % for HSC2 cells and 92 % for HSC4 cells. ",
"Similarly pretreatment with the TRPA1 antagonist reduced thermal allodynia by 83 % for HSC2 cells and 76 % for HSC4 cells (Fig.",
" [4](#Fig4){ref-type=\"fig\"}). ",
"The antagonists did not have an effect on the contralateral paws. ",
"To confirm that the effect of antagonist is peripheral and not systemic, we injected AMG517 or HC030031 in contralateral paw and 45 min later tested the effect of extract-induced thermal allodynia in the ipsilateral paw. ",
"Figure [4c](#Fig4){ref-type=\"fig\"} shows that unlike injection of the TRP antagonist in the ipsilateral paw, injection of antagonists in the contralateral hindpaw had no effect on lipid-induced thermal allodynia. ",
"These data indicate that the drug acts locally at the site of injection. ",
"The effect of lipids released from HSC3 were not further tested, as the lipid extracts did not produce substantial thermal allodynia. ",
"Interestingly, neither the TRPV1 nor the TRPA1 antagonists reversed lipid-induced mechanical allodynia for any of the three cell lines (Fig.",
" [5](#Fig5){ref-type=\"fig\"}). ",
"Taken together, these data suggests that lipids released by OSCC cells induce thermal allodynia by regulating peripheral TRPV1 and TRPA1 activities but these channels do not mediate lipid-induced peripheral mechanical allodynia.",
"Fig. ",
"4Effect of TRP antagonists on Lipid-induced Thermal Allodynia. ",
"TRPV1 antagonist, AMG517 (100 ug) or TRPA1 antagonist HC030031 (100 ug) was injected in the ipsilateral rat hindpaw ipl 45 min prior to lipid extract injection. ",
"Thermal threshold were measured 10 min post extract injection of the ipsilateral and contralateral paws. **",
"a** 50ul of re-suspended lipid extract made from HSC2 cells was injected. **",
"b** 50 ul of re-suspended lipid extract made from HSC4 cells was injected. **",
"c** AMG517(100ug) or HC030031(100ug) was injected in the contralateral rat hindpaw ipl and 45 min later, HSC2 lipid extract was injected in the ipsilateral paw. ",
"Thermal thresholds were measured 10 min post extract injection of ipsilateral and contralateral paws. ",
"N = 6--8 /group. ",
"Vehicle group was injected with 50ul of 10%DMSO/90 % Mineral Oil. ",
"BL = Baseline thresholds. ",
"Data are represented as mean for each group and error bars indicate SEM. ",
"Data were analyzed using two-way ANOVA with Sidak's post-hoc test with p \\<0.05. ",
"Asterisks indicate \\**p* \\< 0.05; \\*\\**p* \\< 0.01; \\*\\*\\**p* \\< 0.001 and \\*\\*\\*\\**p* \\< 0.0001. ",
"Average cell numbers per well at the time of harvest of conditioned media are indicated. ",
"Arrows indicate the side where vehicle or drug was injectedFig. ",
"5Effect of TRP antagonists on Lipid-induced Mechanical Allodynia. ",
"TRPV1 antagonist, AMG517 (100 ug) or TRPA1 antagonist HC030031 (100 ug) was injected in the rat hindpaw ipl 45 min prior to lipid extract injection. ",
"Thermal threshold were measured 10 min post extract injection of the ipsilateral and contralateral paws. **",
"a** 50ul of re-suspended lipid extract made from HSC2 cells was injected. **",
"b** 50ul of re-suspended lipid extract made from HSC3 cells was injected. **",
"c** 50ul of re-suspended lipid extract made from HSC4 cells was injected. ",
"N = 6--8 /group. ",
"Vehicle group was injected with 50ul of 10%DMSO/90 % Mineral Oil. ",
"BL = Baseline thresholds. ",
"Data are represented as mean for each group and error bars indicate SEM. ",
"Data were analyzed using two-way ANOVA with Sidak's post-hoc test with p \\<0.05. ",
"Asterisks indicate \\**p* \\< 0.05; \\*\\**p* \\< 0.01; \\*\\*\\**p* \\< 0.001 and \\*\\*\\*\\**p* \\< 0.0001. ",
"Average cell numbers per well at the time of harvest of conditioned media are indicated\n\nDiscussion {#Sec13}\n==========\n\nTwo distinct features of pain reports from oral cancer patients are that: 1) the pain is specific to malignant lesions and is not observed in pre-cancerous or non-cancerous lesions \\[[@CR37], [@CR38]\\]; and 2) significant pain occurs even when the tumor is still quite small in size. ",
"These aspects of oral cancer pain are consistent with the hypothesis that oral squamous cell carcinoma cells directly modulate activities of peripheral nociceptors. ",
"The TRP channels are one class of receptors expressed on the membranes of trigeminal primary afferents terminating in the oral cavity \\[[@CR8]--[@CR10], [@CR21]\\]. ",
"Recent reports have shown that TRPV1 may play a role in oral cancer pain. ",
"For example, TRPV1 expression in trigeminal ganglia neurons increases after tumor growth in the oral cavity of rats \\[[@CR7]\\] and ablation of TRPV1^+^ neurons reduced OSCC-induced thermal hyperalgesia \\[[@CR11]\\]. ",
"The current study therefore explored the mechanism by which TRPV1 activities are regulated in peripheral sensory neurons during oral cancer pain. ",
"Polyunsaturated fatty acids (PUFAs) are a possible class of factors since human saliva from oral cancer patients has been reported to have elevated levels of certain TRP-active PUFAs (e.g., linoleic acid, arachidonic acid, HODE, and HETE), compared to normal control patients \\[[@CR20]\\], and many of these PUFAs activate TRPV1 \\[[@CR12]--[@CR19]\\] or TRPA1 \\[[@CR12], [@CR22]--[@CR24]\\]. ",
"We therefore evaluated the role of OSCC-secreted lipids in mediating TRP-dependent oral cancer pain using a combined approach of *in vitro* OSCC cell cultures to generate lipid extracts followed by *in vivo* measurement of nociception in rats. ",
"This approach focuses on direct effects of tumor cells on peripheral nociceptors without involvement of carcinogenesis, activation of the immune system or alterations in the extracellular matrix.",
"\n\nIt has been reported that most HNC patients reporting pain have cancerous lesion in the mouth region \\[[@CR39]\\]. ",
"We therefore, used three human OSCC cell lines for our study, namely, HSC2, HSC3 and HSC4, that were derived from tumors obtained from patients with the primary site of tumor being either the floor of the mouth or the tongue \\[[@CR28]\\].",
"\n\nOur data demonstrate that lipids extracted from conditioned media of the OSCC cell lines evoked nocifensive behavior as well as significant thermal and mechanical allodynia (Figs.",
" [1a](#Fig1){ref-type=\"fig\"}, [2b, c, and d](#Fig2){ref-type=\"fig\"} and [3a, b, and c](#Fig3){ref-type=\"fig\"}). ",
"Interestingly, the responses observed for each of the three behavioral modalities, were different between the three cell lines. ",
"This finding of OSCC heterogeneity may contribute to the observed high variation in pain intensity reported by HNC patients. ",
"Importantly, these nociceptive substances appear to be derived from cancerous cells since there were no nociceptive responses observed with lipid extracts from normal oral keratinocyte cells. ",
"Moreover, the collection of conditioned media precluded any contribution from other cell types (e.g., immune cells, endothelium, etc.) ",
"that might co-exist in a tumor mass grown in vivo. ",
"Thus, OSCC release bioactive substances that modulate nociceptor activity. ",
"Prior studies have reported that OSCC cells may release certain protein or peptides such as endothelin-1 \\[[@CR40]\\], nerve growth factor (NGF) \\[[@CR41]\\] or trypsin \\[[@CR42]\\], to activate peripheral afferents. ",
"However, these and other hydrophilic compounds are excluded by the Folch's extraction method \\[[@CR29]\\] and thus the present findings demonstrate, for the first time, that lipids released from OSCC cells may contribute to oral cancer pain.",
"\n\nThe data also demonstrate a key role for TRPV1 in mediating nociception induced by OSCC-derived lipid extracts. ",
"Pre-treatment with a TRPV1 antagonist completely abolished lipid-mediated nocifensive responses and significantly inhibited thermal allodynia. ",
"Further, the data indicate that the site of action of the TRPV1 antagonist was peripheral since injection of the drug in the contralateral paw did not affect lipid-induced thermal allodynia. ",
"However, TRPV1 is not the only receptor system contributing to lipid-induced nociception. ",
"This conclusion is based on two findings. ",
"First, a TRPA1 antagonist had significant attenuation of thermal allodynia. ",
"Second, neither the TRPV1 nor the TRPA1 antagonist altered mechanical allodynia due to OSCC-derived lipids. ",
"Thus, OSCC-released lipids evoke nociception via multiple mechanisms of action. ",
"Identifying the specific lipids from OSCC cells can provide further insights into mechanisms by which these lipids induce oral cancer pain as well as, ways to target their release for development of novel analgesics to treat oral cancer pain.",
"\n\n**Competing interest**\n\nThe authors declare that they have no competing interests.",
"\n\n**Authors' contributions**\n\nSR developed the hypothesis, designed the experiments and analyzed the data. ",
"MB along with DR performed all the behavior assays. ",
"AW performed tissue culture and prepared lipid extracts. ",
"All authors have read and approved the manuscript.",
"\n\nThe study was supported by UTHSCSA seed funding (to SR) and funding obtained from William and Ella Owen\\'s Medical Research Foundation (to SR).",
"\n"
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"Developing a scale to measure parental worry and their attitudes toward childhood cancer after successful completion of treatment: a pilot study.",
"\nThe purpose of this study is to develop and evaluate the psychometric characteristics of a scale designed to measure parents' attitude toward childhood cancer after treatment has ended. ",
"In this study, the 2 theoretical frameworks (theory of attitude and theory of worry) were used as they related to the study. ",
"An attitude is an idea charged with negative or positive emotion, directed to a psychological object, such as cancer. ",
"A sample of 84 White, middle class, American parents (n = 49 mothers, n = 35 fathers) of 51 childhood cancer survivors, whose treatment ended between 1988 and 2005, was surveyed between November 2005 and February 2006. ",
"Two factors were extracted using principal component analysis with oblique rotation. ",
"Cronbach's alpha reliability for Factor 1 was .91 and for Factor 2 was .76. ",
"This study suggests that most parents of cancer survivors tend to perseverate, ruminating on the idea that their child's cancer will return; as a result, they remain in a heightened state of alertness and develop uncontrollable thoughts, or inconsolable worry, about the recurrence of the disease."
] | {
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"Cyberknife radiosurgery for cranial plasma cell tumor.",
"\nCranial and intracranial involvement by myelomatous disease is relatively uncommon. ",
"Furthermore, systemic manifestations of multiple myeloma are present in the majority of these cases at the time of symptom onset. ",
"The authors report the case of a patient with serial appearance of multiple intracranial plasma cell tumor localizations as the first manifestations of a multiple myeloma. ",
"The patient was treated with CyberKnife radiosurgery for a lesion localized at the clivus and sella turcica with complete local control. ",
"With such a technique, based on high-dose conformality, the tumor was centered with an ablative dose of radiation and, at the same time, with a low dose spreading to the surrounding critical structures. ",
"The radiosensitivity of plasma cell tumors renders this treatment modality particularly advantageous for their localized manifestation. ",
"A technical description of this case is provided. ",
"To our knowledge, this is the first case of successful Cyberknife radiosurgery of multifocal intracranial plasmacytoma."
] | {
"pile_set_name": "PubMed Abstracts"
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[
"Value Engineering\n\nRelated links\n\nWhat is Value Engineering?",
"\n\nQuestions?",
"\n\nIf you would like more information about MnDOT's VE program, have a project that meets the FHWA requirements, or would like to be on a VE team, please contact Anjani (Minnie) Milkert, State Value Engineer at minnie.milkert@state.mn.us or 651-366-4648.",
"\n\nIn accordance with the Federal Highway Administration Federal-aid Highway program, MnDOT is required to establish a program to assure that Value Engineering studies are performed on all federal-aid highway projects; and that all resulting, approved recommendations are incorporated into the plans, specifications and project estimate.",
"\n\nThe FHWA defines VE as:\n\n\"The systematic application of recognized techniques by a multi-disciplined team to identify the function of a product or service, establish a worth for that function, generate alternatives through the use of creative thinking, and provide the needed functions to accomplish the original purpose of the project, reliably, and at the lowest life-cycle cost without sacrificing safety, necessary quality, and environmental attributes of the project. ",
"The successful application of the VE process can contribute measurable benefits to the quality of the surface transportation improvement projects and to the effective delivery of the overall Federal-Aid Highway Program.\"",
"\n\nFederal regulations\n\nFederal Regulation 23 CFR Part 627--Value Engineering, requires that states apply VE to all federal-aid highway projects on the National Highway System (NHS) with an estimated total project cost of $25 million or more. ",
"In 2005, an additional requirement was added to include VE studies for bridge projects with an estimated total cost of $20 million or more.",
"\n\nProjects are defined as \"a portion of a highway that a state proposes to construct, reconstruct, or improve as described in the preliminary design report or applicable environmental document\". ",
"A project may consist of several contracts or phases over several years and applies to Design-Build projects as well.",
"\n\nProject estimated cost\n\nThe FHWA can withhold Federal-aid highway funds on any eligible project that did not receive a VE study. ",
"It is strongly recommended that projects approaching the above project regulatory thresholds have a VE study done. ",
"It is likely that project costs will creep up beyond the threshold amount once all totals are in. ",
"If this occurs at the end of the project, it may be too late to conduct a viable VE study.",
"\n\nFunding eligibility\n\nThe cost of performing VE studies is project related and is, therefore, eligible for reimbursement with Federal-aid highway funds at the appropriate pro-rata share for the project studied.",
"\n\nProject managers' responsibility\n\nProject Managers must ensure that a VE Study is performed on all projects meeting the Federal requirements. ",
"Project Managers may also consider a VE Study on lower cost, unique projects that they feel could benefit from a VE study.",
"\n\nProject Managers are responsible for informing the State Value Engineer of all VE recommendations that were implemented and their cost savings, no later than September 30 of each year.",
"\n\nIt is essential that the VE study begin as soon as the Draft Environmental Documents are close to being completed. ",
"VE studies take a considerable amount of time to arrange; therefore, early notification and coordination are essential so that sound VE recommendations can be implemented without delaying the progress of the project. ",
"As a general guideline, allow 6 months to arrange for the VE study.",
"\n\nUse of consultant VE facilitators\n\nMnDOT contracts with qualified VE consultants to conduct the VE studies. ",
"Consulting firms cannot facilitate VE studies on their own designs without an adequate internal firewall.",
"\n\nFacilitators must be qualified VE practitioners, experienced in performing and leading VE studies (have participated in several VE studies as a team member and as a team leader), and have sufficient VE training, education, and experience to be recognized by SAVE International as meeting the requirements for certification. ",
"Please refer to the Pre-Qualification Program Overview webpage for more information.",
"\n\nUse of consultant VE team members\n\nMnDOT does contract with qualified consultants (with specific expertise) for the role of VE team member. ",
"Consulting VE team members cannot participate on VE study teams when their firm had involvement with the design project under study.",
"\n\nDesign-build projects\n\nIn the case of Federal-aid design-build projects meeting the above project cost threshold criteria, a VE study must be performed prior to the release of the Request for Proposals document.",
"\n\nState value engineer’s responsibility\n\nThe Office of Project Management & Technical Support currently manages MnDOT’s VE program. ",
"The State Value Engineer is responsible for documenting the results of VE studies (both workshop and change proposals) for inclusion in the annual VE status report that is submitted to the FHWA at the end of the Federal Fiscal year (September 30). ",
"The State Value Engineer provides all necessary VE related assistance to the Project Managers."
] | {
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[
"What happens when two supermassive black holes collide? ",
"Combining the observing power of two future ESA missions, Athena and LISA, would allow us to study these cosmic clashes and their mysterious aftermath for the first ...\n\nAstronomy\n\nMay 23, 2019\n\n1\n\n171\n\nSupermassive black hole\n\nA supermassive black hole is a black hole with a mass of the order of between 105 and 1010 solar masses. ",
"Most, if not all, galaxies, including the Milky Way, are believed to contain supermassive black holes at their centers.",
"\n\nSupermassive black holes have properties which distinguish them from their relatively low-mass cousins:\n\nYour Privacy\n\nThis site uses cookies to assist with navigation, analyse your use of our services, and provide content from third parties.",
"\nBy using our site, you acknowledge that you have read and understand our Privacy Policy\nand Terms of Use."
] | {
"pile_set_name": "Pile-CC"
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"Culturing of avian embryos for time-lapse imaging.",
"\nMonitoring morphogenetic processes, at high resolution over time, has been a long-standing goal of many developmental cell biologists. ",
"It is critical to image cells in their natural environment whenever possible; however, imaging many warm-blooded vertebrates, especially mammals, is problematic. ",
"At early stages of development, birds are ideal for imaging, since the avian body plan is very similar to that of mammals. ",
"We have devised a culturing technique that allows for the acquisition of high-resolution differential interference contrast and epifluorescence images of developing avian embryos in a 4-D (3-D + time) system. ",
"The resulting information, from intact embryos, is derived from an area encompassing several millimeters, at micrometer resolution for up to 30 h."
] | {
"pile_set_name": "PubMed Abstracts"
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0,
0,
0,
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[
"Q:\n\nDynamic width animation based in CSS Media Query\n\nI have two divs:\n<div class=\"map-ctn\"></div>\n<div class=\"list-ctn\"></div>\n\nAnd the Media Queries:\n@media only screen and (min-width: 1281px) and (max-width : 1824px) {\n .map-ctn { width: 80%; }\n .list-ctn { width: 20%; }\n}\n\n@media only screen and (min-width: 961px) and (max-width : 1280px) \n{\n .map-ctn { width: 75%; }\n .list-ctn { width: 25%; }\n}\n\nI want to start the .list-ctn div with 0% and the .map-ctn with 100% and animate the .list-ctn do X% and .map-ctn to Z% based on the Media Query active.",
"\nEDIT:\nThe animation should occur on a jQuery event (form submit).",
"\nSo maybe I need to control the divs using jQuery but using the Média Query info to animate the width based on the width of each div.",
"\nIt has to work from IE9+, Safari, Chrome and Firefox.",
"\n\nA:\n\nsetTimeout(function() {\r\n $('.mapexpanded').removeClass('mapexpanded');\r\n}, 300);\nAnd the Media Queries:\r\n@media only screen and (min-width: 1281px) and (max-width : 1824px) {\r\n .map-ctn { width: 80%; }\r\n .list-ctn { width: 20%; }\r\n}\r\n\r\n@media only screen and (min-width: 961px) and (max-width : 1280px) \r\n{\r\n .map-ctn { width: 75%; }\r\n .list-ctn { width: 25%; }\r\n}\r\n\r\n.map-ctn { width: 60%; }\r\n.list-ctn {width: 40%; }\r\n\r\n.map-ctn, .list-ctn {\r\n transition: width 1s ease-in-out;\r\n height: 300px;\r\n background-color: red;\r\n margin-bottom: 10px;\r\n}\r\n\r\nhtml body .mapexpanded .map-ctn {\r\n width: 100%;\r\n}\r\nhtml body .mapexpanded .list-ctn {\r\n width: 0%;\r\n}\n<script src=\"https://ajax.googleapis.com/ajax/libs/jquery/2.1.1/jquery.min.js\"></script>\r\n<div class=\"mapexpanded\">\r\n <div class=\"map-ctn zerowidth\"></div>\r\n <div class=\"list-ctn zerowidth\"></div>\r\n</div>\n\nI used JQuery here only to shorten the JS a bit; not because it makes heavy use of any JQuery libraries. ",
"The animation going on here is all CSS.",
"\nYou didn't have a width for viewports that didn't match your queries, so I just added 60%/40%. ",
"It's possible you actually wanted something else; feel free to tweak it. ",
"It will shift automatically when the browser is resized. ",
"Also, you can likely shorten the setTimeout call, or replace it with requestAnimationFrame.",
"\n\n"
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[
"/*\n * Tencent is pleased to support the open source community by making BK-CI 蓝鲸持续集成平台 available.",
"\n *\n * Copyright (C) 2019 THL A29 Limited, a Tencent company. ",
" All rights reserved.",
"\n *\n * BK-CI 蓝鲸持续集成平台 is licensed under the MIT license.",
"\n *\n * A copy of the MIT License is included in this file.",
"\n *\n *\n * Terms of the MIT License:\n * ---------------------------------------------------\n * Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation\n * files (the \"Software\"), to deal in the Software without restriction, including without limitation the rights to use, copy,\n * modify, merge, publish, distribute, sublicense, and/or sell copies of the Software, and to permit persons to whom the\n * Software is furnished to do so, subject to the following conditions:\n *\n * The above copyright notice and this permission notice shall be included in all copies or substantial portions of the Software.",
"\n *\n * THE SOFTWARE IS PROVIDED \"AS IS\", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR IMPLIED, INCLUDING BUT NOT\n * LIMITED TO THE WARRANTIES OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. ",
"IN\n * NO EVENT SHALL THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER LIABILITY,\n * WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, OUT OF OR IN CONNECTION WITH THE\n * SOFTWARE OR THE USE OR OTHER DEALINGS IN THE SOFTWARE.",
"\n */\n\npackage com.tencent.devops.websocket.controller\n\nimport com.tencent.devops.websocket.pojo.",
"ChangePageDTO\nimport com.tencent.devops.websocket.pojo.",
"ClearUserDTO\nimport com.tencent.devops.websocket.pojo.",
"LoginOutDTO\nimport com.tencent.devops.websocket.servcie.",
"WebsocketService\nimport org.springframework.beans.factory.annotation.",
"Autowired\nimport org.springframework.messaging.handler.annotation.",
"MessageMapping\nimport org.springframework.stereotype.",
"Controller\n\n@Controller\nclass WsController @Autowired constructor(\n val websocketService: WebsocketService\n) {\n\n @MessageMapping(\"/changePage\")\n fun changePage(changePage: ChangePageDTO) {\n websocketService.changePage(changePage.userId, changePage.sessionId, changePage.page, changePage.projectId, changePage.showProjectList)\n }\n\n @MessageMapping(\"/loginOut\")\n fun loginOut(loginOutDTO: LoginOutDTO) {\n websocketService.loginOut(loginOutDTO.userId, loginOutDTO.sessionId, loginOutDTO.page, loginOutDTO.transferData)\n }\n\n @MessageMapping(\"/clearUserSession\")\n fun clearUserSession(clearUserDTO: ClearUserDTO) {\n websocketService.clearUserSession(clearUserDTO.userId, clearUserDTO.sessionId, clearUserDTO.transferData)\n }\n}"
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"// WARNING: DO NOT EDIT THIS FILE. ",
"THIS FILE IS MANAGED BY SPRING ROO.",
"\n// You may push code into the target .java compilation unit if you wish to edit any member(s).",
"\n\npackage nl.bzk.brp.model.data.kern;\n\nimport nl.bzk.brp.model.data.kern.",
"HisPersgeslachtsaand;\nimport org.springframework.beans.factory.annotation.",
"Configurable;\n\nprivileged aspect HisPersgeslachtsaand_Roo_Configurable {\n \n declare @type: HisPersgeslachtsaand: @Configurable;\n \n}\n"
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"mu-Protocadherin, a novel developmentally regulated protocadherin with mucin-like domains.",
"\nBranching morphogenesis is a central event during the development of kidneys, lungs, and other organs. ",
"We previously generated a monoclonal antibody, 3D2-E9, that inhibited branching morphogenesis and caused widespread apoptosis. ",
"We now report the purification of its antigen and cloning of its full-length cDNA. ",
"Its cDNA encodes an integral membrane protein that contains four cadherin-like ectodomains and a thrice tandemly repeated region enriched in threonine, serine, and proline, similar to those of mucins. ",
"We thus term this protein mu-protocadherin, reflecting the hybrid nature of its extracellular region. ",
"mu-Protocadherin is expressed in two forms that are developmentally regulated, with the shorter isoform lacking the mucin-like repeats. ",
"Expression of the long isoform in heterologous cells results in adhesion of the expressing cells, suggesting that it is a new cell adhesion molecule. ",
"mu-Protocadherin contains both N and O glycosylations. ",
"It is expressed at lateral and basal surfaces of epithelia during kidney and lung development and is located in coated pits. ",
"Colocalization of mu-protocadherin with beta-catenin was noted primarily at the junction of the lateral and basal membrane. ",
"The cytoplasmic domain contains four proline-rich regions, similar to SH3 binding regions. ",
"Thus, it is likely that adhesive interactions mediated by mu-protocadherin induce signaling events that regulate branching morphogenesis."
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"GamePolitics - Comments for \"&#039;Saints Row Penetrator&#039; Altered In Saints Row The Third for Japan\"http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan\nComments for \"'Saints Row Penetrator' Altered In Saints Row The Third for Japan\"enRe: 'Saints Row Penetrator' Altered In Saints Row The Third ...http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan#comment-275668\n<p>That comment just made my day.</p>\nMon, 14 Nov 2011 18:55:35 +0000ddrfr33kcomment 275668 at http://gamepolitics.comRe: 'Saints Row Penetrator' Altered In Saints Row The Third ...http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan#comment-275653\n<p>Actually, no, Rapelay not okay. ",
"It was hit by the political wave that originated from here. ",
"Now a huge amount of similar game, also including it, have stricter regulation, if not directly a ban, can't remember.</p>\nSun, 13 Nov 2011 20:13:59 +0000DorthLouscomment 275653 at http://gamepolitics.comRe: 'Saints Row Penetrator' Altered In Saints Row The Third ...http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan#comment-275646\n<p>Tentacle porn okay. ",
"Rapelay okay. ",
"Purple dildo on a stick bad.<img alt=\"laugh\" height=\"20\" src=\"http://gamepolitics.com/sites/all/libraries/ckeditor/plugins/smiley/images/teeth_smile.gif\" title=\"laugh\" width=\"20\" /></p>\nSat, 12 Nov 2011 14:46:26 +0000Thomas Riordancomment 275646 at http://gamepolitics.comRe: 'Saints Row Penetrator' Altered In Saints Row The Third ...http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan#comment-275645\n<p>I guess it just ain't that graphic without the ding dongs and hoo hahs.</p>\n<p> </p>\n<p>Andrew Eisen</p>\nSat, 12 Nov 2011 04:45:31 +0000Andrew Eisencomment 275645 at http://gamepolitics.comRe: 'Saints Row Penetrator' Altered In Saints Row The Third ...http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan#comment-275642\n<p>I wish Japan would get rid of the "no genitals" thing. &",
"nbsp;It's led to something that seems truly schizophrenic in my mind. &",
"nbsp;It's ok to show a graphic depiction of rape, just so long as ding dongs and hoo hahs are censored. &",
"nbsp;The mind boggles.</p>\nSat, 12 Nov 2011 02:44:42 +0000Davidcomment 275642 at http://gamepolitics.comRe: 'Saints Row Penetrator' Altered In Saints Row The Third ...http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan#comment-275636\n<p>A three-foot long purple rubber dildo attached to the handle of a baseball bat is not a genital part. So what's the problem?</p>\n<p> </p>\n<p>Andrew Eisen</p>\nSat, 12 Nov 2011 00:35:00 +0000Andrew Eisencomment 275636 at http://gamepolitics.comRe: 'Saints Row Penetrator' Altered In Saints Row The Third ...http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan#comment-275631\n<p>the censorship forces them to be more "creative", otherwise it'd be close-up penetration shots like in the US. ",
"it's also worth noting that sextoys are not censored in porn there; I'm guessing that they can't be shown with great detail in non-pornographic work</p>\nFri, 11 Nov 2011 21:42:35 +0000Kincyrcomment 275631 at http://gamepolitics.comRe: 'Saints Row Penetrator' Altered In Saints Row The Third ...http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan#comment-275626\nWhat a wasted opportunity to do a tentacle variant.",
"Fri, 11 Nov 2011 20:23:11 +0000Cattleprodcomment 275626 at http://gamepolitics.comRe: 'Saints Row Penetrator' Altered In Saints Row The Third ...http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan#comment-275624\n<p>They should have just pixelated it, would have been funnier.</p>\nFri, 11 Nov 2011 20:20:42 +0000hellfire7885comment 275624 at http://gamepolitics.comRe: 'Saints Row Penetrator' Altered In Saints Row The Third ...http://gamepolitics.com/2011/11/11/039saints-row-penetrator039-altered-saints-row-third-japan#comment-275621\n<p>""Depiction of genital parts" is prohibited in Japan."</p>\n<p>And yet, paradoxically, Japan manages to have the most depraved and diverse selection of pornography the world has ever seen.</p>\nFri, 11 Nov 2011 19:20:31 +0000Zerodashcomment 275621 at http://gamepolitics.com"
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[
"Q:\n\nLogin plugin of TinyAuth for CakePHP 3\n\nI have a problem with the login of the user, I have different tables for each user because they have different atributes (students, administrators, profesors) and I don't know how can I configure TinyAuth for more than one UserModel. ",
"Sorry for my poor english. ",
"Thx\n\nA:\n\nTo make your life easier you should keep them all under the same users table, but have a groups table as well. ",
"The groups table will have students, professors and administrators. ",
"The users table will in turn have a group_id column where each user can have a reference to the groups table. ",
"\n|users|\n-> id\n-> group_ip\n-> username\n-> first_name\n-> last_name\n-> created\n-> updated\n\n|groups|\n-> id\n-> name\n-> created\n-> updated\n\nUsing this technique requires you to have the right relations setup.",
"\nIf you don't like to use groups, you can have a role field in your users table. ",
"\n|users|\n-> id\n-> role\n-> username\n-> first_name\n-> last_name\n-> created\n-> updated\n\nRegardless of the choice you pick, you should not create multiple tables and expect uniform login to work nicely. ",
"\nYou should follow CakePHP's Blog tutorial to have a better understanding of how the role technique is used.",
"\nhttp://book.cakephp.org/3.0/en/tutorials-and-examples/blog-auth-example/auth.html\n\n"
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"Disassembly of micelles in nanoscopic space to prepare concentric nanotubes with variable hydrophobic interiors.",
"\nNanotubes with variable hydrophobic interiors were prepared by disassembling anionic micelles in the inner walls of positive charge bearing conjugated polymer nanotubes."
] | {
"pile_set_name": "PubMed Abstracts"
} | [
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[
"This directory contains test cases for individual source features of LLVM.",
"\nIt is designed to make sure that the major components of LLVM support all of the\nfeatures of LLVM, for very small examples. ",
" Entire programs should not go here.",
"\n\nRegression tests for individual bug fixes should go into the test/Regression dir.",
"\n\n"
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"Jake Anderson\n\nDallasTexas\n\nFull-time technology blogger, love to travel and spend time with friends over the weekends.",
"\n\nFavorite movie dialog: \"Live long, and prosper.\"",
"\n\nAwestruck by Star Trek as a kid, Jake has been relentless in his pursuit for covering the big technological innovations that will shape the future. ",
"A self-proclaimed gadget freak, he loves getting his hands on every piece of gadget that comes his way."
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[
"\"Archer.\" \"",
"Lana?\" \"",
"Lana!\" \"",
"Whoa, Jesus Christ!\" \"",
"Chill out, Miggs!\" \"",
"I'm sorry, it's just been so long.\" \"",
"Okay, it's been...\" \"Oh, my God.\" \"",
"You're pregnant again?\" \"!\" \"",
"What?\" \"",
"How long have I been down here, left to rot in this godforsaken hole?\" \"!\" \"",
"Like... three weeks.\" \"",
"Hmm, seems so much longer than that.\" \"",
"It...\" \"So, please tell me that's an entire canard a la presse with a file in it.\" \"",
"Half a chicken, no file, just that weird quasi-cornbread they have here.\" \"",
"Ugh, why do you even bother?\" \"",
"That's weird, I was asking myself the same thing on the way down here.\" \"",
"And then brought crappy food anyway?\" \"",
"And then left?\" \"",
"No, no, no, wait, come on, Lana, I-I'm sorry, but\" \"I-I'm starting to lose it down here.\" \"",
"You-you got to make him let me go!\" \"",
"Make him how?\" \"",
"Well, I hate to even ask, but...\" \"Blow him?\" \"",
"Your words.\" \"",
"And then what, Archer, you want me to blow the whole army?\" \"",
"How is there still an army?\" \"",
"How have the rebels not wiped them out already?\" \"",
"Because turns out, he's actually very good at commanding an army.\" \"",
"And as Third and\" \"Fourth Battalion withdraw, here comes First Armored, and now the rebels are outflanked.\" \"",
"COMANDANTESir, it's brilliant!\" \"",
"A classic pincher movement.\" \"",
"Just like Hannibal at the battle of Cannae.\" \"",
"And also Keith, in my\" \"Warhammer guild.\" \"",
"So, get used to that weird cornbread, 'cause you might be down here a...\" \"Lana, I need a woman's touch!\" \"",
"Oh!\" \"",
"Ow!\" \"",
"I'm sorry, Lana.\" \"",
"I said a woman, not a stevedore who lost his hand in a stevedoring accident and then got a hand transplant from an actual bear.\" \"",
"So, did you want to just do it right through the bars or...?\" \"",
"I mean, we do.\" \"",
"And sometimes they do.\" \"",
"Hello.\" \"",
"I... wait.\" \"",
"So, in three weeks, you go from you're gonna execute Archer to now you guys are, like, swinger pals?\" \"",
"Well, I wouldn't say \"pals.\"\" \"",
"Wow, not what you said yesterday.\" \"",
"Oh, come on, that was the pruno talking.\" \"",
"Well, I hate to break up this delightful ménage, but\" \"Presidente Cyril wants her back upstairs.\" \"",
"Wait, what?\" \"",
"What?\" \"",
"Why.\" \"",
"Guess.\" \"",
"Ah.\" \"",
"Mm.\" \"",
"Ow!\" \"",
"Oh, wait, wait, wait, wait, wait, wait, wait!\" \"",
"What?\" \"",
"Archer, do you want me to move over one?\" \"",
"Eh.\" \"",
"Ah, Madame First Lady.\" \"",
"So good of you to join me.\" \"",
"It's not as if I had a choice.\" \"",
"Well...\" \"Nor am I still the the first lady.\" \"",
"Well, maybe not technically, since your husband divorced you and then was immediately deposed by me, but the situation is, as they say, fluid.\" \"",
"Ugh.\" \"",
"I brought her, can I go now?\" \"",
"Yes, for the love of\" \"God, get out.\" \"",
"I meant home.\" \"",
"PAMWhatareyou , nuts?\" \"",
"Cap'n Crunch here just took all the coca fields back from the rebels.\" \"",
"We're sitting on an endless supply.\" \"",
"Of what, coca leaf smoothies?\" \"",
"Oh, my God.\" \"",
"That's a coca leaf smoothie.\" \"",
"And they're gross.\" \"",
"It basically tastes like a fart of itself, but it's better than the cocaine donuts and whipped cocaine she was eating.\" \"",
"'Cause that's just empty calories.\" \"",
"Well, and cocaine.\" \"",
"Which, if I'm being totally honest, I maybe might have a problem with.\" \"",
"No.\" \"",
"And, Lana, the rebels still control the airport.\" \"",
"It's their HQ.\" \"",
"So, unless you want to walk, you can't go home.\" \"",
"And why on Earth would you want to?\" \"",
"Uh...\" \"Why bother going back?\" \"",
"The U.S. government stole your livelihood, forced you to become a criminal.\" \"",
"And I hate to say it, but it's not like you have a man waiting for you.\" \"",
"I...\" \"You know...\" \"Whereas here we have everything we need.\" \"",
"It's like San Marcos has been waiting thousands of years for us to arrive.\" \"",
"Lana, this could be our...\" \"Don't.\" \"",
"Zion, yes, there, I said it.\" \"",
"Wait, they got weed here, too?\" \"",
"That's not what the song's about!\" \"",
"Told you.\" \"",
"But I just this second realized why you want to stay down here.\" \"",
"Cyril's just the Vader to your\" \"Palpatine.\" \"",
"Is that a reference\" \"I should get, or...?\" \"",
"Seriously?\" \"",
"Well, I'm sorry,\" \"Lana, I didn't go to rabbinical school.\" \"",
"It's from...\" \"No, you know what?\" \"",
"Never mind.\" \"",
"But, spoiler alert, Vader ends up killing Palpatine.\" \"",
"Hey.\" \"",
"But only for three days, right?\" \"",
"And then he moves the rock and comes out of his cave stronger than ever.\" \"",
"Then he shuts off the tractor beam, releasing the Jews from captivity.\" \"",
"What?\" \"",
"Hey.\" \"",
"Wait, yes, now that sounds familiar.\" \"",
"It cannot possibly.\" \"",
"Gospel of Luke?\" \"",
"Hey.\" \"",
"Oh for...\" \"Ray, you used to be a preacher, you want to back me up here?\" \"",
"I actually don't know.\" \"",
"My church didn't really do the\" \"New Testament.\" \"",
"The one with Jesus\" \"Christ in it.\" \"",
"I mean, I take your word for it, but...\" \"Hey!\" \"",
"Will you idiots get out?\" \"!\" \"",
"I'm trying to talk to the first lady!\" \"",
"Of what, this country or country music?\" \"",
"'Cause I'm first lady of both!\" \"",
"CYRIL No, you're not...\" \"Outlaw country!\" \"",
"Oh, my God, my beautiful and also multi-talented wife!\" \"",
"You are here!\" \"",
"I know, right?\" \"",
"It's like, finally.\" \"",
"As in... this is a good thing?\" \"",
"Duh!\" \"",
"I'm finally gonna get the outlaw country cred I deserve.\" \"",
"Oh, my God, you've always deserved it.\" \"",
"Wait, how does being in jail...?\" \"",
"Paging Dr. Black,\" \"Dr. Man In Black!\" \"",
"Johnny Cash was in Folsom Prison and San Quentin, so...\" \"Yeah, to visit.\" \"",
"Hmm?\" \"",
"He just played for the prisoners, dummy.\" \"",
"He wasn't actually in prison.\" \"",
"But don't worry.\" \"",
"We're gonna get out of here and go join the rebels.\" \"",
"Yes, then you and\" \"I, my beautiful wife, will live a simple life in the hills, scratching shit food, like yams, out of the ground with a, uh, yam-scratcher.\" \"",
"Okay, A... start counting backwards from a million years, and when you get to zero that still won't happen.\" \"",
"Aw.\" \"",
"And B... how... how do we get out of here?\" \"",
"Well, we're, uh, still fine-tuning it, but...\" \"We have no plan, whatsoever.\" \"",
"Okay, good, because\" \"I didn't want to step on anybody's toes, but byoink.\" \"",
"Whaaaaat?\" \"",
"Where did you get that?\" \"",
"Let's just say,\" \"\"I honestly don't know.\"\" \"",
"Fair enough.\" \"",
"Cyril says the rebel headquarters are...\" \"Wait, is it \"are\" or is it \"is\"?\" \"",
"It's \"where,\" idiot!\" \"",
"The airport!\" \"",
"Jesus!\" \"",
"And you, sir, will keep a civil tongue in your head when you are addressing the First Lady of the government-in-exile of La\" \"República de San Marcos!\" \"",
"And also country music.\" \"",
"I facetiously beg milady's pardon.\" \"",
"Oh, milady facetiously grants it.\" \"",
"Oh, goody.\" \"",
"Now let's steal a car, go join the rebels, then come back here and crush Cyril's stupid army.\" \"",
"You make it sound so easy.\" \"",
"ARCHERWell, it's Cyril.\" \"",
"I mean, unless he's got a nuclear warhead in his pants...\" \"Which he totally does.\" \"",
"Which, guys, can we talk about this?\" \"",
"The time for talk is kaput, Klon-Bruder.\" \"",
"Now is the time for action!\" \"",
"Clone bone!\" \"",
"And by that I mean his... you know...\" \"Yeah, we...\" \"Let me finish!\" \"...",
"penis.\" \"",
"No, you ruined it.\" \"",
"And by \"it,\" I meant the moment.\" \"",
"Not his penis.\" \"",
"I...\" \"Penis.\" \"",
"Will you shut up?\" \"!\" \"",
"I know what I'm doing!\" \"",
"Yes, but don't do what you are doing!\" \"",
"Are you crazy?\" \"!\" \"",
"This car is worth one quarter of\" \"$1 million!\" \"",
"Well, was.\" \"",
"And did you ever think maybe your fleet of priceless cars is the reason you're fighting off a rebellion?\" \"",
"I'm fighting the rebels because that is what we do!\" \"",
"My father fought the rebels, his father fought the rebels...\" \"So, like a family business.\" \"",
"That manufactures oppression.\" \"",
"Well, and cocaine.\" \"",
"In fact...\" \"Stop!\" \"",
"What?\" \"!\" \"",
"That.\" \"",
"I mean, if you think we have time.\" \"",
"I...\" \"Is there a tiger?\" \"",
"Of course there is a tiger.\" \"",
"Where did you think you are,\" \"Tegucigalpa?\" \"",
"And so, by the power self-vested in me, I now pronounce us man and wife, and I may also kiss the bride.\" \"",
"Yay!\" \"",
"Ugh, second worst wedding ever.\" \"",
"Yeah, but at least nobody died.\" \"",
"PAMYet!\" \"",
"Inappropes.\" \"",
"What is wrong with you?\" \"",
"I'm trying to make that a thing people say, like,\" \"\"Damn, dawg, that's inappropes!\"\" \"",
"Plus, I think I'm addicted to cocaine.\" \"",
"Well, but look on the bright side.\" \"",
"With any luck at all, it will save me the trouble of killing you myself.\" \"",
"PAMDamn, dawg...\" \"Inappropes.\" \"",
"Yay!\" \"",
"ARCHERYay!\" \"",
"Yay!\" \"",
"They should've sent a poet.\" \"",
"Oh, and some meat.\" \"",
"Do we have any meat?\" \"",
"Literally tons.\" \"",
"No, I didn't mean another animal.\" \"",
"I meant like a steak, or some...\" \"Typical American.\" \"",
"You think meat comes from the supermercado, all wrapped in a nice plastic wrap.\" \"",
"Yeah, you're describing meat.\" \"",
"No, no, no.\" \"",
"Meat is blood and bones and sinew!\" \"",
"Well, now you're describing not-meat.\" \"",
"Meat is whatever the tiger says is meat, because\" \"God made him the boss and all the other animals his food!\" \"",
"Hang on, buddy.\" \"",
"And thank you, George Bore-well, for that clunky analogy in defense of totalitarianism, but\" \"I'm not feeding Shane an elephant.\" \"",
"Who the hell is...\" \"Oh, my God, he's totally a\" \"Shane.\" \"",
"So let me bounce this off you guys...\" \"What say we don't launch this giant nerve gas missile at, uh... where did you say you were launching it?\" \"",
"Wouldn't you like to know?\" \"",
"I would, actually.\" \"",
"Come on, you guys are acting a little... insane.\" \"",
"Do you know what's insane?\" \"!\" \"",
"Yeah, this.\" \"",
"All of it, everything.\" \"",
"The rocket, uh...\" \"I'm not really comfortable with the clone-bone.\" \"",
"You're not doing it right.\" \"",
"Now move, so I can begin the launch sequence.\" \"",
"Over my dead body, man that was a jinx, huh?\" \"",
"No, no, no, wait, wait, wait!\" \"",
"Hang on!\" \"",
"As amazing as it would be, we can't take Shane with us.\" \"",
"Yeah, we can, watch.\" \"",
"No, no, no, no!\" \"",
"No, don't!\" \"",
"Okay, okay, okay, okay.\" \"",
"Carol, listen to me very closely.\" \"",
"Why, you gonna walk me through your organic method of vaginal hygiene?\" \"",
"My what?\" \"",
"You're a bigger pussy than Shane!\" \"",
"Come here, boy!\" \"",
"Shane!\" \"",
"Do something!\" \"",
"Like what?\" \"",
"Get him!\" \"",
"We're coming, brother!\" \"",
"Hiy...!\" \"",
"No...!\" \"",
"Brother...!\" \"",
"You bastard!\" \"",
"Brother, the launch sequence!\" \"",
"No!\" \"",
"Don't stop!\" \"",
"Almost... there.\" \"",
"Oh, come on!\" \"",
"Shut... up.\" \"",
"Launch sequence initiated.\" \"",
"No!\" \"",
"KRIEGERCLONE1No!\" \"",
"Ha-ha!\" \"",
"It doesn't work with a bow... tiiiiiiiiieeeeee!\" \"",
"Shane!\" \"",
"Shane...!\" \"",
"Shane...!\" \"",
"Shane?\" \"",
"Well, we should've guessed that was gonna happen.\" \"",
"W... we should've?\" \"",
"Well, or at least could've.\" \"",
"I mean...\" \"Yeah, I-I-I guess, uh...\" \"I guess a leopard can't change his spots.\" \"",
"Uh...\" \"I don't know that much about leopards.\" \"",
"That was actually a proverb.\" \"",
"That was actually a tiger.\" \"",
"Minutes to launch: 180.\" \"",
"Huh.\" \"",
"That is a lot longer than I thought it would be.\" \"",
"Okay, when we get to rebel HQ, they might be aggressive at first, but...\" \"They wouldn't if we had a damn tiger.\" \"",
"No.\" \"",
"We're-we're not going back.\" \"",
"Unless...\" \"No.\" \"",
"What if...\" \"He had his chance!\" \"",
"A what?\" \"",
"A charivari.\" \"",
"Also called a shivaree?\" \"",
"You know, it's that thing where everybody bangs pots and pans outside our room as we... consummate.\" \"",
"Oh... n-no.\" \"",
"We're not doing that.\" \"",
"Ha-ha!\" \"",
"Ow!\" \"",
"Ray, my feet are already killing me.\" \"",
"Sorry.\" \"",
"He's probably not used to leading.\" \"",
"You know...\" \"You, on the other hand...\" \"lead away.\" \"",
"Mind if I cut in?\" \"",
"No.\" \"",
"Yes!\" \"",
"Screw you guys.\" \"",
"Hey, Krieger!\" \"",
"What the hell is going on?\" \"",
"Cyril's the new dictator...\" \"President.\" \"",
"Shut up, and he just married crazy tits, and I need a dance partner.\" \"",
"So nobody noticed that I've been missing for three weeks?\" \"",
"Um...\" \"Well, I certainly didn't.\" \"",
"We figured the vampires\" \"The what?\" \"",
"Wait, what?\" \"",
"KRIEGERThey'renot vampires, they're my clones.\" \"",
"Or I'm their clone, or...\" \"Oh, please, you and your clone nonsense, it's all a bunch of...\" \"Holy shit.\" \"",
"Why was I not informed of this, Krieger?\" \"",
"If he even is Krieger!\" \"",
"Huh?\" \"",
"The real Krieger could be down there!\" \"",
"Oh, for... that is just...\" \"I won't even dignify that with a response.\" \"",
"Uh, uh...\" \"Pam?\" \"",
"Pam!\" \"",
"Uh-huh.\" \"",
"I've got a concussion.\" \"",
"And this regular tie means nothing, I don't like bow ties so I switched.\" \"",
"Well, you certainly sound concussed.\" \"",
"He sounds like a vampire!\" \"",
"I say we tie him up and let the sun sort it out!\" \"",
"You need to hush.\" \"",
"Minutes to launch: 150.\" \"",
"What is that?\" \"",
"What is going on?\" \"",
"Tell me that robot's not talking about launching this rocket, Krieger.\" \"",
"Or should we say\" \"Count Krieg...\" \"Ow!\" \"",
"Ow!\" \"",
"Ow, cut it out!\" \"",
"Ow!\" \"",
"Goddamn it!\" \"",
"Ow!\" \"",
"Thank you.\" \"",
"Vampire lovers.\" \"",
"Yeah, so, here's the thing...\" \"This is it, rebel headquarters.\" \"",
"Now, remember, let me do the talking.\" \"",
"Got it.\" \"",
"I'm serious.\" \"",
"I got it.\" \"",
"Look at me.\" \"",
"I got it!\" \"",
"Jesus!\" \"",
"Okay.\" \"",
"Then here we go.\" \"",
"Pinche cabrón!\" \"",
"Ow!\" \"",
"Goddamn it!\" \"",
"You said you'd let me do the... !\" \"",
"Espere, espere!\" \"!\" \"",
"Mira, mira!\" \"",
"Es la cantante!\" \"",
"It's all about me this time...\" \"Aw...\" \"So.\" \"",
"You're saying this gigantic rocket is not only full of deadly nerve gas, but also aimed\" \"God knows where?\" \"",
"Well, that statement presupposes the existence of\" \"God, but...\" \"Krieger, I am about two seconds away from telling\" \"Pam to get a wooden stake.\" \"",
"Yay!\" \"",
"I'm not a vampire!\" \"",
"Doesn't matter to the stake.\" \"",
"Then yes!\" \"",
"It's full of nerve gas, and I don't know where it's aimed!\" \"",
"What?\" \"",
"What's the range on this thing?\" \"",
"I don't know, 2,000 miles?\" \"",
"How far away is the\" \"Upper East Side?\" \"",
"I don't know, 1,980-something miles?\" \"",
"Okay, this is a presidential order from the president.\" \"",
"Shut it down!\" \"",
"I can't!\" \"",
"Can't, or won't?\" \"",
"Can't!\" \"",
"Oh.\" \"",
"I didn't hear you.\" \"",
"Ray, can you override the launch sequence?\" \"",
"Um...\" \"Launch sequence accelerated.\" \"",
"No.\" \"",
"Oh, for God's sake.\" \"",
"Minutes to launch: 90.\" \"",
"Cyril...\" \"Uh, it's actually\" \"Senor Presidente, but...\" \"Shh, hush.\" \"",
"Now I know you've had a fun time as dictator, and we all enjoyed this little break from him, but Cyril... and if you tell him\" \"I said this, I'll break your frickin' arms... it's... ugh...\" \"Archer time.\" \"",
"Wha...?\" \"",
"Oh, all right.\" \"",
"Krieger, I assume the whatever...\" \"Doppelkriegers.\" \"",
"I assume they had a lab, so take Ray and see if you can find some info on how to shut this damn thing down.\" \"",
"We'll go get Archer.\" \"",
"Ooh, and Cherlene!\" \"",
"Ooh, and no.\" \"",
"Because she isn't here.\" \"",
"Oh!\" \"",
"Damn it, Archer!\" \"",
"Damn you!\" \"",
"You can't even let me pretend to be magnanimous while inwardly gloating!\" \"",
"Oh, shut up.\" \"",
"What does the note say?\" \"",
"LANA\" \"So, airport?\" \"",
"But the note says not...\" \"Well, I guess that settles that!\" \"",
"I mean, it's not like we can just waltz into enemy territory and...\" \"Well, I certainly can't.\" \"",
"Can't, or...\" \"Both, either, all.\" \"",
"They'd shoot me on sight, Lana.\" \"",
"And if you want Archer so damn bad, you can go get him your...\" \"Huh.\" \"",
"I didn't think she'd actually do it.\" \"",
"Then you're an idiot.\" \"",
"Oh, I'm the idiot?\" \"",
"Yes.\" \"",
"What the...?\" \"",
"Holly?\" \"",
"Archer?\" \"",
"Rando?\" \"",
"Slater?\" \"",
"Who the hell is Rando?\" \"",
"Him!\" \"",
"That's Sterling\" \"Archer!\" \"",
"CHERLENEA-hem!\" \"",
"And I am Cherlene?\" \"",
"The Queen of Outlaw Country?\" \"",
"What the hell are you doing here?\" \"",
"What the hell are you doing here?\" \"",
"I could ask you the same question.\" \"",
"I amasking you the same question!\" \"",
"We came to join the rebels.\" \"",
"What?\" \"",
"What's the FBI doing in San Marcos?\" \"",
"With a cocaine smuggler?\" \"",
"I'm not FBI, and he's not a...\" \"Well, technically,\" \"I am, but...\" \"We're both CIA.\" \"",
"What?\" \"!\" \"",
"CHERLENEC ...\" \"I...\" \"A.\" \"And we're also the rebels.\" \"",
"And you are in a mundo of caca.\" \"",
"Okay!\" \"",
"Listen, stop, listen to me!\" \"",
"Lana?\" \"!\" \"",
"Holly?\" \"",
"What's the FBI doing...\" \"These dickheads aren't FBI, Lana, they're CIA!\" \"",
"And also the rebels!\" \"",
"Huh.\" \"",
"Thought that would get more of a reaction.\" \"",
"Yeah, no, that is, um... news, uh... but my water just broke, so...\" \"Captioned by Media Access Group at WGBH access.wgbh.org\" \"Made... in Georgia.\""
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[
"The effects of ryanodine on calcium uptake by the sarcoplasmic reticulum of ischemic and reperfused rat myocardium.",
"\nThe effects of ischemia and reperfusion on sarcoplasmic reticulum (SR) calcium uptake were measured in crude heart homogenates of rats and were compared to published results for rabbit hearts. ",
"Isolated rat hearts (n = 5 in each group) were Langendorff-perfused at 37 degrees C and were either kept normally perfused (control group), or submitted to 15 min normothermic ischemia (ischemic group), or reperfused for 10 min after 15 min ischemia (reperfused group). ",
"Mechanical function recovered to 50-60% of control after 10 min reperfusion following ischemia. ",
"Ca uptake (control Vmax: 23.0 +/- 2.20 nmol.min.1.mg of protein-1) decreased during ischemia (Vmax: 15.7 +/- 1.60 nmol.min-1.mg-1) but recovered to control level on reperfusion (Vmax: 20.8 +/- 2.02 nmol.min-1.mg-1). ",
"An increased Ca uptake was obtained when the measurements were carried out in the presence of ryanodine (430 microM) to block Ca leakage through SR Ca-release channels. ",
"The relative magnitude of ryanodine effect in the ischemic myocardium (increase: 77.2 +/- 18.20%) was more marked than in control (32.0 +/- 8.22%) or reperfused myocardium (39.0 +/- 10.66%). ",
"This result is different from that of rabbit myocardium where similar ryanodine effect is present in all groups (56.7 +/- 13.76%, 50.0 +/- 13.56% and 54.2 +/- 6.88% in control, ischemic and reperfused hearts, respectively) and suggests that a component of cytosolic Ca overload via SR Ca-release channels is present during ischemia in rat, but not in rabbit myocardium."
] | {
"pile_set_name": "PubMed Abstracts"
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[
"/*\n * The Alluxio Open Foundation licenses this work under the Apache License, version 2.0\n * (the \"License\"). ",
"You may not use this work except in compliance with the License, which is\n * available at www.apache.org/licenses/LICENSE-2.0\n *\n * This software is distributed on an \"AS IS\" basis, WITHOUT WARRANTIES OR CONDITIONS OF ANY KIND,\n * either express or implied, as more fully set forth in the License.",
"\n *\n * See the NOTICE file distributed with this work for information regarding copyright ownership.",
"\n */\n\npackage alluxio.wire;\n\nimport com.google.common.base.",
"MoreObjects;\nimport com.google.common.base.",
"Objects;\n\nimport javax.annotation.concurrent.",
"NotThreadSafe;\n\n/**\n * Class that represents the available, free, and total capacity.",
"\n */\n@NotThreadSafe\npublic class Capacity {\n private long mTotal;\n private long mUsed;\n\n /**\n * Creates a new instance of {@link Capacity}.",
"\n */\n public Capacity() {}\n\n /**\n * @return the total capacity\n */\n public long getTotal() {\n return mTotal;\n }\n\n /**\n * @return the used capacity\n */\n public long getUsed() {\n return mUsed;\n }\n\n /**\n * @param total the total capacity to use\n * @return the capacity\n */\n public Capacity setTotal(long total) {\n mTotal = total;\n return this;\n }\n\n /**\n * @param used the used capacity to use\n * @return the capacity\n */\n public Capacity setUsed(long used) {\n mUsed = used;\n return this;\n }\n\n @Override\n public boolean equals(Object o) {\n if (this == o) {\n return true;\n }\n if (!(",
"o instanceof Capacity)) {\n return false;\n }\n Capacity that = (Capacity) o;\n return mTotal == that.mTotal && mUsed == that.mUsed;\n }\n\n @Override\n public int hashCode() {\n return Objects.hashCode(mTotal, mUsed);\n }\n\n @Override\n public String toString() {\n return MoreObjects.toStringHelper(this).add(\"total\", mTotal).add(\"used\", mUsed).toString();\n }\n}\n"
] | {
"pile_set_name": "Github"
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[
"--source include/have_32bit.inc\n--source suite/sys_vars/inc/parser_max_mem_size_basic.inc\n"
] | {
"pile_set_name": "Github"
} | [
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] | 0 | 5 |
[
"Shari Hoort\n\nIt is always a great thing, and sometimes a rare thing, when one can say that they love their job! ",
"I am very privileged to have been given the opportunity to say that.",
"\n\nAt Simply Counted Business Services, I am able to do what I love on a daily basis. ",
"I also get to learn and expand my knowledge. ",
"I enjoy working with numbers and helping our clients keep their records in order.",
"\n\nThe atmosphere in this office is amazing, everyone is so ready to jump in and help each other. ",
"This extends to the way we treat our clients, we care about each of them and want each one to feel important.",
"\n\nI graduated from Calvin College with a BA in Business Administration. ",
"I have 20 years of bookkeeping experience in a local real estate/property management office. ",
"I loved it there and thought that I would be there forever, but the economy changed.",
"\n\nI have chosen to move in a different direction for the last few years. ",
"In addition to working at Simply Counted, I am a school bus driver for West Ottawa Public Schools. ",
"They say diversity is good!",
"\n\nI met my husband while at Calvin College. ",
"We have 2 great sons, 1 amazing daughter-in-law and 2 delightful grandsons.",
"\n\nIn my spare time, I enjoy camping, kayaking, spending time with family and being involved with church. ",
"I have also been a mentor in the Kids Hope Program."
] | {
"pile_set_name": "Pile-CC"
} | [
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[
"Q:\n\nAdd missing data to a dictionary in python\n\nMy dictionary represents data from 24 hours. ",
"The dictionary fills dynamically depending of the hour. ",
"If at some hour there was no data, then it skips that hour. ",
"Please. ",
"\nI would like to add the keys (which represent hours) that are not in the dictionary and give them a value of 0. ",
"The dictionary below is how I get the data: \n{\n '0': 1, \n '1': 1, \n '4': 1, \n '7': 1, \n '8': 4, \n '9': 6, \n '10': 3, \n '11': 12, \n '12': 11, \n '13': 9, \n '14': 8, \n '15': 7, \n '16': 5, \n '17': 9, \n '18': 9, \n '19': 5, \n '20': 7, \n '21': 4, \n '22': 2, \n '23': 4\n}\n\nThis is how I would like to have my final dictionary: \n{\n '0': 1, \n '1': 1,\n '2': 0,\n '3': 0,\n '4': 1, \n '5': 0,\n '6': 0,\n '7': 1, \n '8': 4, \n '9': 6, \n '10': 3, \n '11': 12, \n '12': 11, \n '13': 9, \n '14': 8, \n '15': 7, \n '16': 5, \n '17': 9, \n '18': 9, \n '19': 5, \n '20': 7, \n '21': 4, \n '22': 2, \n '23': 4\n}\n\nThanks!",
"\n\nA:\n\nUse dict.get() in a for loop:\nmy_dict = {'0': 1, '1': 1, '4': 1, '7': 1, '8': 4, '9': 6, '10': 3, '11': 12, '12': 11, '13': 9, '14': 8, '15': 7, '16': 5, '17': 9, '18': 9, '19': 5, '20': 7, '21': 4, '22': 2, '23': 4}\n\nmy_dict = {str(i): my_dict.get(str(i), 0) for i in range(24)}\n\n"
] | {
"pile_set_name": "StackExchange"
} | [
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[
"Exposure-response relationships for work-related neck and shoulder musculoskeletal disorders--Analyses of pooled uniform data sets.",
"\nThere is a lack of quantitative data regarding exposure-response relationships between occupational risk factors and musculoskeletal disorders in the neck and shoulders. ",
"We explored such relationships in pooled data from a series of our cross-sectional studies. ",
"We recorded the prevalence of complaints/discomfort (Nordic Questionnaire) and diagnoses (physical examination) in 33 groups (24 female and 9 male) within which the workers had similar work tasks (3141 workers, of which 817 were males). ",
"In representative sub-groups, we recorded postures and velocities of the head (N = 299) and right upper arm (inclinometry; N = 306), right wrist postures and velocities (electrogoniometry; N = 499), and muscular activity (electromyography) in the right trapezius muscle (N = 431) and forearm extensors (N = 206). ",
"We also assessed the psychosocial work environment (Job Content Questionnaire). ",
"Uni- and multivariate linear meta-regression analysis revealed several statistically significant group-wise associations. ",
"Neck disorders were associated with head inclination, upper arm elevation, muscle activity of the trapezius and forearm extensors and wrist posture and angular velocity. ",
"Right-side shoulder disorders were associated with head and upper arm velocity, activity in the trapezius and forearm extensor muscles and wrist posture and angular velocity. ",
"The psychosocial work environment (low job control, job strain and isostrain) was also associated with disorders. ",
"Women exhibited a higher prevalence of neck and shoulder complaints and tension neck syndrome than men, when adjusting for postures, velocities, muscular activity or psychosocial exposure. ",
"In conclusion, the analyses established quantitative exposure-response relationships between neck and shoulder disorders and objective measures of the physical workload on the arm. ",
"Such information can be used for risk assessment in different occupations/work tasks, to establish quantitative exposure limits, and for the evaluation of preventive measures."
] | {
"pile_set_name": "PubMed Abstracts"
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[
"We used to keep Sheep,\nOn one occasion we had an Ewe with a prolapsed uterus (the uterus had dropped down below the cervix after birth of Lamb).",
"\nIt was sticking out of the rear end, like a red footbal.",
"\nWe got the Vet in, I had to hold the Sheep upside down, whilst the vet pushed & shoved to get it all back in. (",
"like a postman trying to deliver a pillow through a letterbox).",
"\nThe Vet, then stiched her Fanny up, by using a \"Duffle bag Stitch\" which he then drew tight.",
"\nAfter a while, the Uterus dropped again, but couldnt fully pop-out because of the stitching.",
"\nSo we had this Ewe waliking round like John Wayne\nBleating in an embarrassed fashion, with an enormous lump at the rear end, with a stiched up Fanny on the outer edge of the Lump.",
"\n.......\n\nSinnick, quelle drollerie about the hurting ass! ",
"Le Vrai Rikkor has a great sense of dignity. ",
"Even if I try to put a basebell cap on his head (and he would do anything for me), he moves away from the situation quickly. ",
"Some dogs are smart enough to know when they are being made sport of.",
"\n\nMy dog used to cry when it left the park - every time. ",
"I had to explain to people why she wasn't budging and why she was making that awful sobbing noise. ",
"Similar to the noise you make when you lose a derby match - yeah it was that bad and that loud."
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"Story highlights \"Legitimate rebellion\" staged in Valencia, quickly contained\n\nGovernment official calls the operation a \"terrorist attack of a paramilitary nature\"\n\nCaracas, Venezuela (CNN) Venezuela remained a powder keg on Sunday as authorities said they had quelled an anti-government paramilitary attack at a military base that led to the deaths of two people, and the country's attorney general defied her ouster by the newly elected National Constituent Assembly.",
"\n\nA man who identified himself as an army officer announced the revolt on social media, an action he called a \"legitimate rebellion\" aimed at the government of leftist President Nicolás Maduro.",
"\n\n\"We are united now, more than ever, with the brave people of Venezuela who do not recognize the legitimacy of Nicolás Maduro's murderous tyranny,\" according to a man who said he was Capt. ",
"Juan Caguaripano.",
"\n\nThe President, speaking on his weekly TV show, \"Sundays with Maduro,\" made reference to the incident, saying \"a week ago, we won with votes and today we had to beat terrorism with bullets.\"",
"\n\n\"They attack with terrorism and hate. ",
"We attack with our work, our love. ",
"They destruct, we construct,\" he said.",
"\n\nRead More"
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[
"We reward effective, well-managed local organisations for their past work, shining a deserving spotlight on their success and providing support that lets them steer, sustain or scale their work on their own terms. ",
"http://www.starsfoundation.org.uk/\n\nGram Vikas founders came to Orissa in the early 1970s as student volunteers to serve victims of a devastating cyclone. ",
"Their extensive activism and relief work motivated them to form Gram Vikas, which was registered on January 22, 1979, and currently serves more than 3,89,333 people in 1196 habitations of 25 districts in Odisha. ",
"Through its direct outreach programmes Gram Vikas works in 943 villages across 23 districts covering 59,132 families of which 39% are adivasis, 14% are dalits and the remainder are from general castes, mostly poor and marginal farmers.",
"\n\nMobile Creches works with birth to 12 year old children living on the construction sites and slums of Delhi(NCR) since 1969. ",
"Integrated Daycare Centers for children on construction sites are run. ",
"Mobile Creches has so far reached out to 7,50,000 children, trained 6,500 women as childcare workers, run 650 daycare centres and partnered with 200 builders.",
"\n\nMobile Creches runs daycares centres where children are provided education, nutrition and healthcare. ",
"Community-women and NGOs are trained to provide care and speak-up for the child. ",
"The years of experience along with strong-networks helps in the advocacy for policy-change.",
"\n\nWe reward effective, well-managed local organisations for their past work, shining a deserving spotlight on their success and providing support that lets them steer, sustain or scale their work on their own terms. ",
"http://www.starsfoundation.org.uk/\n\nWe reward effective, well-managed local organisations for their past work, shining a deserving spotlight on their success and providing support that lets them steer, sustain or scale their work on their own terms. ",
"http://www.starsfoundation.org.uk/"
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[
"USL Players Help New Zealand Hold Peru\n\nWELLINGTON, New Zealand – Vancouver Whitecaps FC 2’s Deklan Wynne and former Reno 1868 FC loanee Kip Colvey both went the full 90 minutes as New Zealand earned a 0-0 draw with Peru in the opening leg of the intercontinental playoff between the countries that will decide one of the final places at the 2018 FIFA World Cup.",
"\n\nA sellout crowd of more than 37,000 fans at Westpac Stadium in Wellington saw New Zealand almost grab a late winner through Ryan Thomas, whose long-range shot clipped the outside of the post, but the underdog All Whites more than held their own against the visitors, who recorded three shots on goal by were unable to find a way past Vancouver Whitecaps FC goalkeeper Stefan Marinovic.",
"\n\nWynne, who made 22 appearances for WFC2 this past season, and Colvey, who made 10 appearances on loan with 1868 FC in its inaugural season, were joined on the field in the second half by Portland Timbers 2 midfielder Bill Tuiloma, who came on with 11 minutes to go.",
"\n\nThe second leg between New Zealand and Peru is set for Wednesday night at the Estadio Nacional in Lima, Peru, with kickoff at 9:15 p.m. ET."
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[
"Hey there! ",
"So I'm writing this as I read, and everything will most likely read disconnected and not have any fluency to it at all.",
"\n\nI like the opening passages and the sorrow which you so beautifully convey, but I have to admit it's kind of confusing. ",
"When you refer to people by only their appearance, it gets really confusing to keep up with who's who, and I think that often, only one initial appearance description is enough to suffice. ",
"When you overuse it, it, again, gets confusing and turns people off immediately (believe me, I've been told-off for it many a time XD)\n\nI really liked the description of Marlene and how she affected the friendships of the group. ",
"In so little words you created an entire character who I can see, know, relate to. ",
"I think it's because you described her as a friend would describe her, not as a narrator would, and that voice put more life into her than a narrator could.",
"\n\nShe lifted it to eye level, toasting herself. ",
"She was throwing a pity party, one invite only. ",
"Drink and cry, wash away the tears, drown the sadness. ",
"Life was a bitch, why not drink to her health?",
"\n\nMost likely the most brilliant line I've ever read, where pity parties are concerned. ",
"To this point the most prominent thing I'm noticing is how consistent the changes in emotions are. ",
"Nothing is abrupt, as sadness usually goes (one second they're sad, the next they're angry, the next their weeping uncontrollably...), but everything really flows together so that the reader is feeling the emotions with the character, and not just getting slapped in the face with them. ",
"To add, as I've--as has most of the world--been in that place where a noisy bar and a good beer is all you want, I think you really... portrayed that well. ",
"As bars are the most common settings I've seen in the fictions I read, I think you've really played on all the right tones (explaining why she went there, her treatment to the bartender, the quoted line).",
"\n\nAnother thing that I do often:\n\n“Insult you?” ",
"snarled Sirius, his temper finally flaring\n\nYou slipped PoV, or perspective, or whatever. ",
"The way you've written it, it describes his emotion as he or a narrator would describe it, not Lily. ",
"As we're seeing and hearing Lily's voice, she would have to say something along the lines of \"She could see his temper flaring\" or \"his eyes burned with a finally flaring temper\" or just something that describes it from Lily's point of view. ",
"It's such a small and insubstantial line, but to the nitpicker like me (who's gotten told a hundred times over that I don't keep PoV up) it stands out.",
"\n\nI've found what's been poking me this whole time:\n\nDorcas’s eyes flashed. “",
"I wasn’t offering it,” she snapped.",
"\n\nYou have a really good knack for characterization; not canon-wise, just person-wise. ",
"Attitudes aren't flat and continuous, but they actually change as people's actually do. ",
"I'm not quite sure how to elaborate on that, but it's really refreshing to read.",
"\n\nHalt, it's the spelling police! (",
"Yeah, I'm kind of corny)\n\nDorcas nodded. “",
"I hope you decided to; you need to get out of here.”",
"\n\nSurprised danced across his face at the sight of her, but he grinned easily. “",
"Hey, Lily.”",
"\n\nSo overall I really enjoyed this, but the ending wasn't quite what I expected (I'm not really sure what I expected, so I can't image that's of much help. ",
":D) One thing I could point out though is that there wasn't an... ending, to Sirius and Lily's relationship-- it didn't have any finality. ",
"I know that her going to James is supposed to be that finality, but... I don't know. ",
"To end this lamely, I really enjoyed reading this. :)",
"\n\n-Ari\n\nReviewer: fg_weasley Date: 04/09/09 3:04 Chapter: As the Dawn Breaks\n\nOh, Mere. [",
"hugs]\n\nNow you know my love for Sirius/Lily. ",
"And you know my love of your writing. ",
"So you know I had to come and read this. ",
"And holy cheese and rice am I glad I did. ",
"You make me proud to be a Sirius/Lily shipper, my dear.",
"\n\nFirst, I have to squee a bit at the fact that the prompt that I posted was the inspiration for this masterpiece. ",
"It makes me smile; not just because I posted it, but because its Janet Fitch. ",
"Isn’t it such a beautiful, wonderful, Janet Fitch-y quote? ",
"LOL. ",
"And I had hoped this quote would be used for Sirius/Lily, because that was who I had in mind when I put up that quote. ",
":D You definitely did it justice, dear.",
"\n\nAnyway. ",
"Enough of that and back to your actual story.",
"\n\nThe way you set up the scene at the beginning was great, because it not only set up the picture, but it set up the mood as well. ",
"Having the news of Marlene’s death as its own, solitary paragraph was excellent stylistically. ",
"It gave the news that much force and it was perfect.",
"\n\nI really like that you kept their identities vague at the beginning. ",
"Rather than say Lily, James, and Sirius you kept it much more interesting and showed great charactization, such as here:\n\nThe other, though, kept his face solid and unwavering, completely emotionless. ",
"Once or twice, but no more than that, grief would cloud his eyes for a second. ",
"The only other emotion he showed was a hint of worry as his gaze shifted from his drink on the table to the crying redhead on the other side of his hazel-eyed friend.",
"\n\nI like that part because you show that Sirius doesn’t hide his emotions, per se, but he doesn’t wear them on his sleeve as Lily does. ",
"From this paragraph I just got this utter sense of Sirius. ",
"Its overwhelming, and I think that is why it shows such great charactization. ",
"The reader knows exactly who it is without the author having said it outright. ",
"Well done.",
"\n\nThe firecracker peddler who could distract everyone in an instant if Sirius or Lily said something too intimate or made a wrong gesture that could’ve let their secret out.",
"\n\nI really loved this hint. ",
"It made me smile. ",
":D I also like that it shows that Sirius and Lily trusted her enough to know of their secret. ",
"In my experience of writing and reading this pairing, I don’t think they’ve ever told anyone – it’s always only been between them. ",
"I like the twist you have, and the way you wove in Marlene. ",
"This is a great hint early on to Marlene’s part in the story, which is strengthened later by the flashbacks.",
"\n\nThe characterizations were, I thought, well done. ",
"Little things, like the paragraph I mentioned above, really show how well you know both Lily and Sirius. ",
"This made me laugh: “It’s whatever was in the cupboard.” ",
"He gave her a lopsided smile. “",
"I think I like fuck buddy a lot better—it has a nice ring to it.” ",
"Yet at the same time, I really think it is something Sirius would say, and especially something that would make Lily roll her eyes or not acknowledge at all, which is exactly what you have her do.",
"\n\nAs I said before, you really handled the quote well. ",
"I think you showed both emotions – love and hate – vividly, and I like the way you played with both. ",
"I think the best part, for me, and the best example, is here: “I hate him,” she whispered to the still room. ",
"Something stirred in her as the words broke the haunting quiet. ",
"Encouraged by the feeling, she said it again, louder, “I hate him.” ",
"The raw emotion is there, in that whole scene, and you know your ability to portray emotion is what I love most about your writing in general. ",
"You did not fail here.",
"\n\nThe end, of course, I simultaneously liked and disliked. ",
"I disliked it only because I’m biased and partial to Sirius/Lily and James is a prat. ",
"Lol. ",
"However, getting down to the actual writing and the story, I liked it. ",
"It was a great way to show Lily and James ultimately together, thus adhering to canon, yet still showing a bit of a connection between Sirius and Lily. ",
"After all, one thing love and hate have in common is deep feeling. ",
"But I also liked that you didn’t make it seem like she was only going with James because Sirius told her she should; I like to believe that, even though I dislike the pair, Lily went with James because she truly loved him. ",
"And with your end you showed the beginnings of that. ",
"Well done, dear.",
"\n\nTwo things:\n\nSirius sighed and rand his hands through his hair.",
"\n\nShould be ran, not rand.",
"\n\nLily rolled her eyes, the thudding in her head starting up again—the bassist had founded the unplugged cord.",
"\n\nFound, instead of founded? ",
"Also, while I’m here, I liked the metaphor of the band in her head – nice touch.",
"\n\nOh, Mere. ",
"This was very, very good. ",
"Very real, very evocative. ",
"Angsty, but not excessively so, with a hopeful ending and a few light touches to balance it.",
"\n\nYou seem to have a gift for characterisation. ",
"Even Marlene, without really appearing in the fic, has a personality - caring and mischievous. ",
"She just leaps off the screen. ",
"Lily's emotions when she's in her flat before going to the bar are very realistic to me, too, her grief and utter desperation. ",
"You really made me feel what she was feeling. ",
"Later on, as well, when she's feigning being okay and almost babbling - that was an excellent touch, and would be humourous if it wasn't all a cover for Lily's immense pain. ",
"James - oh, I felt sorry for him, poor sweet naive James. ",
"You were true to his character: trusting in his best friend and so in love with Lily that he didn't notice anything between them. ",
"And Sirius... well, you managed to avoid making him a total cliche, while still having him as a seducer. ",
"Well done! ",
"He was complex, too: I wasn't quite sure whether to be charmed or repulsed. ",
"The latter, I think. ",
"The way he seemed respectful to Marlene but then seemed to be not just celebrating her memory, but actually taking advantage of the Lily's state to seduce her... he wastes no time, and it seems to be a calculated move, not one fueled by alcohol, which makes him quite a jerk, really. ",
"And then his claims of having to end things with her because he loved her - well, it's impossible to know how truthful he is there, hmm...\n\nYou come up with fantastic metaphors, as well. ",
"I loved the band/headache one. ",
"However, I must say it seemed a bit awkward having that play such a prominent role in one passage of the fic (especially as it's quite Muggle-ish) but not appearing anywhere else - it could have been really effective if you'd woven it in earlier and had music as a theme tying the whole fic together. ",
"Anyway, I also adored this passage:\n\nDetermination, anger, and hatred surged through her, powering her up out of her chair and into the shower. ",
"She washed off a week of sweat, tears, and heartbreak. ",
"She scrubbed away the drowning grief of Marlene’s murder. ",
"She rinsed off the life-sucking power of Sirius’s sugar-coated, I-love-you break up.",
"\n\nJust... *loves*. ",
"The repetition of 'she' makes it cohesive, but the different washing-related words give it variation... wow, it's just really well-written and empowering.",
"\n\nYou captured the passion of Sirius and Lily's encounter very well without being graphic, too.",
"\n\nUnfortunately, there were a few spots where I thought the writing could be more polished - things could be clearer and more eloquently expressed. ",
"For example, in the opening passage, I think you overused descriptions of eyes to refer to the characters. ",
"And in the very first passage, 'group' and 'seemed' kept appearing - I don't think the last sentence really needs to be there, actually. ",
"Perhaps you need to focus a bit on keeping your writing simple - your expressiveness and wonderful plot and characterisation will still be there even when you cut out a few words.",
"\n\nI loved how you used the quote and the pairing and came up with this fantastic unique fic that puts James, Lily and Sirius into new lights - and fits with canon! ",
"You explored the fine line dividing love and hatred, and I like that in the end Sirius' behaviour provided the means of Lily coming to appreciate James.",
"\n\nAll in all, Mere, your storytelling is fantastic. ",
"This was a well-constructed fic with excellent characterisation and grasp on emotions. ",
"Chipping away just a tiny bit more will make your gems sparkle even more brightly :)\n\nAww! ",
"Mere, you know I loved it, because I love all of your other fics, but I felt soooooooooo unbelieveably bad for Sirius. ",
"Siriusly! (",
"I'm done with the bad puns now...) It was brilliantly written, the emotions were portrayed amazingly, and I can just see this happening in canon. ",
"The grief I feel for Sirius, however, is immense. ",
"But, due to the wonderfulness of the rest of the story overshadowing the grief, I will forgive you. ;)",
"Are you planning to write a companion fic to this? ",
"Like, a sequel, or maybe what Sirius does after Lily throws him out? ",
"Ooh or maybe a prequel! ",
"Haha sorry... It was great, though! {",
"BeccA}\n\nI liked this - I’ve read a few Sirius/Lily fics lately, and this one was quite different. ",
"I really didn’t expect their relationship to just, well, end. ",
"I can see why Sirius left, but I can also see it from Lily’s point of view.",
"\n\nThe way you compared Lily’s headache from the drinking to a band thumping in her head was good - and I liked how you kept this idea consistent throughout that scene. ",
"And, I know Lily blows her top eventually, but the first time she sniped at him, I didn’t see why she didn’t follow it through - push the point.",
"\n\nThe quote in the beginning was really well chosen - it fit with your story like a glove. ",
"And the end was nice - I like how you made it fit with canon like that, and after a rather angsty story, her going to James and getting a new flatmate showed that there was a way forward.",
"\n\nShe stood up, like a spring uncoiling.",
"\n\nI love that line - the imagery is really quite strong for something so simple.",
"\n\nNit-picks:\n\n“I’m serious, Lily.” ",
"She could practically feel him willing her to take him seriously. - ",
"you repeat ‘serious’ and that made me stop.",
"\n\nShe dropped her purse on her only table in the kitchen, a tube of lipstick and a pen rolling out. - ",
"I’d suggest a semi-colon instead of a comma, and ‘rolled’. ",
"It would make more sense, I think. ",
"Also, ‘pen’ is rather Muggle. ",
"I know Lily is Muggle-born, but I wouldn’t imagine her taking a pen to the Hog’s Head.",
"\n\nShe swallowed again shaking her head, but it didn’t help. - ",
"a comma after ‘again’ would work well, I think.",
"\n\nNo! ",
"her mind yelled. ",
"No! ",
"Not possible. ",
"Marly wouldn’t—couldn’t die. - ",
"I think you meant to italicise ‘no’ and possibly a hyphen after ‘couldn’t’?",
"\n\nthe long bar that stretched the back wall. - ‘",
"stretched along/across’?",
"\n\nSirius’s eyes hardened as he gave the barman a hard look. - ",
"it would be better without the repetition of ‘hard’.",
"\n\nSirius sighed and rand his hands through his hair. - ‘",
"ran’.",
"\n\nthe bassist had founded the unplugged cord. - ",
"do you mean ‘found’?",
"\n\nShe lifted her head, eyes glistening, and looked at Dorcas’s tender pair. - ",
"after reading this line a couple of times I got it, but at first I didn’t realise ‘tender pair’ meant eyes. ",
"To clear it up ‘into’ might work better than ‘at’. ",
"Not sure.",
"\n\n“But I thought you would want some time,” she said smiling a little. - ",
"a comma after ‘said’.",
"\n\nShe turned at walked down the hall - ‘and’ and not ‘at’.",
"\n\nSurprised danced across his face at the sight of her - ‘surprise’.",
"\n\nAlso, you mentioned a fridge at one point, but also a pantry. ",
"I don’t know much about what they provide you in flats, etc, but to me a fridge seems rather Muggle. ",
"It is electrical, after all. ",
"I probably wouldn’t have noticed that though if it weren’t for the FFTF discussion. ",
"XD"
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[
" IN THE UNITED STATES COURT OF APPEALS\n FOR THE FIFTH CIRCUIT United States Court of Appeals\n Fifth Circuit\n\n FILED\n October 8, 2009\n\n No. ",
"09-30401 Charles R. Fulbruge III\n Summary Calendar Clerk\n\n\n\nAMBER CHAUVIN,\n\n Plaintiff - Appellant\nv.\n\nRADIOSHACK CORP,\n\n Defendant - Appellee\n\n\n\n\n Appeal from the United States District Court\n for the Eastern District of Louisiana\n USDC No. ",
"2:08-CV-4255\n\n\nBefore REAVLEY, JOLLY, and OWEN, Circuit Judges.",
"\nPER CURIAM:*\n The judgment of the district court is affirmed for the following reasons:\n 1. ",
" The jurisdiction of the court existed for the reasons given by the\n order of April 7, 2009. ",
"The plaintiff’s petition appears to predicate\n the alleged wrongful termination on a violation of the plaintiff’s\n entitlement to medical leave of absence. ",
" The claim under the\n federal act was confirmed by plaintiff’s counsel. ",
" Subsequently,\n\n\n *\n Pursuant to 5TH CIR . ",
"R. 47.5, the court has determined that this opinion should not\nbe published and is not precedent except under the limited circumstances set forth in 5TH CIR .",
"\nR. 47.5.4.",
"\n\f No. ",
"09-30401\n Summary Calendar\n\n plaintiff dismissed that claim, but jurisdiction was determined on\n the date of removal.",
"\n 2. ",
" As an at-will employee, plaintiff has no claim for her termination.",
"\n Whether or not she was allowed medical leave is immaterial,\n because that would not change the defendant’s right to terminate\n her employment. ",
"Again, the district court by its order of April 9,\n 2009, correctly ruled.",
"\nAFFIRMED.",
"\n\n\n\n\n 2\n\f"
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[
"Q:\n\nScream very loudly\n\nWrite a script that outputs A to stdout infinitely.",
"\nThere shold be no newlines or seperators between the characters\nStandard loopholes apply\nThis is code-golf. ",
"The shortest solution in each language wins.",
"\n\nA:\n\nx86-16, IBM PC DOS, 7 6 bytes\n00000000: b041 cd29 ebfc .A.)..\n\nUnassembled listing:\nB0 41 MOV AL, 'A' ; put 'A' into AL\n PRINT: \nCD 29 INT 29H ; DOS fast console output char in AL\nEB FC JMP PRINT ; loop infinitely\n\nAs a bonus, if you run this on your IBM 5151 monitor for a few hours this will actually produce infinite output on that screen until the end of time.",
"\n\nA:\n\nTuring Machine Code, 9 bytes\n0 * A r 0\n\nTry it online!",
"\nOne of the very few times that Turing Machine Code can compete overall.",
"\n\nA:\n\nbrainfuck, 16 bytes\n+[+[<]>>+<+]>[.]",
"\n\nCredit to the Brainfuck constants page for 65!",
"\nTry it online!",
"\n\n"
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"This program/project seeks to elucidate the basic mechanisms that regulate the replication and differentiation of both normal and cancer cells. ",
"The ultimate goal is the application of such knowledge to the prevention or treatment of malignant disease in humans. ",
"Significant research progress towards these goals has been made during this past year in these specific project areas: 1. ",
"Studies of the interaction of the transcription machinery with DNA templates using purified RNA polymerases from E. coli and wheat germ and specific cloned fragments of the DNA. ",
"2. ",
"Molecular events in the processing of primary RNA transcripts in nuclei of eukaryotic cells. ",
"3. ",
"Studies of the role of specific DNA sequences in the control of the promotion, termination, and anti-termination of transcription. ",
"4. ",
"The development of genetic systems for studying embryonic development. ",
"5. ",
"The molecular processes involved in the replication and assembly of new chromatin. ",
"6. ",
"The mechanism of action of estrogen and glucocorticoid receptors in the regulation of gene expression. ",
"7. ",
"The role of nuclear and cytoplasmic membranes and the phosphorylation of the constituent membrane proteins in regulation of genetic expression. ",
"8. ",
"Studies of the interaction of tumor-promoting phorbol esters and anti-tumorigenic retinoids in the control of cell replication and differentiation. (",
"V)"
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"Q:\n\nJavaScript: how to filter deep JSON objects\n\nI have an array of deep JSON objects that look like similarly to this:\nvar hierarchy = [\n {\n \"title\": \"category 1\",\n \"children\": [\n {\"title\": \"subcategory 1\",\n \"children\": [\n {\"id\": 1, \"title\": \"name 1\"},\n {\"id\": 2, \"title\": \"name 2\"},\n {\"id\": 3, \"title\": \"name 3\"}\n ]\n },\n {\"title\": \"subcategory 2\",\n \"children\": [\n {\"id\": 1, \"title\": \"name 4\"}\n ]\n }\n ]\n },\n {\n \"title\": \"category 2\",\n \"children\": [etc. - ",
"shortened for brevity]\n }\n];\n\nSo basically it is a hierarchy - there are categories which can have subcategories which contain objects with some IDs and names. ",
"I also have an array of IDs that are related to the deepest hierarchy level (objects with no children) and I need to filter this set of objects in such a way that only (sub)categories that contain defined objects remain.",
"\nSo for example if I had an array containing two IDs:\nvar IDs = [2, 3];\n\nthe result would be: \nvar hierarchy = [\n {\n \"title\": \"category 1\",\n \"children\": [\n {\"title\": \"subcategory 1\",\n \"children\": [\n {\"id\": 2, \"title\": \"name 2\"},\n {\"id\": 3, \"title\": \"name 3\"}\n ]\n }\n ]\n }\n];\n\ni.e. the whole, the whole 'category 2' object removed, the whole 'subcategory 2' removed, object with ID '1' removed.",
"\nThe problem is that the depth of those objects is variable and unknown - some objects have no children, some have children that also have children etc., ",
"any subcategory can can itself have a subcategory and I basically need to find object with no children that have defined IDs and keep the whole path to each of them.",
"\nThank you.",
"\n\nA:\n\nBasically, perform a depth first traversal of your tree invoking a callback function on each node. ",
" If that node is a leaf node and it's ID appears in your list then clone the branch that leads to that leaf, but don't re-clone any part of the branch that was already cloned.",
"\nOnce you have constructed the partial and filtered copy of your tree you need to cleanup the original. ",
" I mutated the original tree in the process for book-keeping purposes - tracking which branches had already been cloned.",
"\nEdit: modified code to filter list of trees instead of just a single tree\nvar currentPath = [];\n\nfunction depthFirstTraversal(o, fn) {\n currentPath.push(o);\n if(o.children) {\n for(var i = 0, len = o.children.length; i < len; i++) {\n depthFirstTraversal(o.children[i], fn);\n }\n }\n fn.call(null, o, currentPath);\n currentPath.pop();\n}\n\nfunction shallowCopy(o) {\n var result = {};\n for(var k in o) {\n if(o.hasOwnProperty(k)) {\n result[k] = o[k];\n }\n }\n return result;\n}\n\nfunction copyNode(node) {\n var n = shallowCopy(node);\n if(n.children) { n.children = []; }\n return n;\n}\n\nfunction filterTree(root, ids) {\n root.copied = copyNode(root); // create a copy of root\n var filteredResult = root.copied;\n\n depthFirstTraversal(root, function(node, branch) {\n // if this is a leaf node _and_ we are looking for its ID\n if( !",
"node.children && ids.indexOf(node.id) !",
"== -1 ) {\n // use the path that the depthFirstTraversal hands us that\n // leads to this leaf. ",
" copy any part of this branch that\n // hasn't been copied, at minimum that will be this leaf\n for(var i = 0, len = branch.length; i < len; i++) {\n if(branch[i].copied) { continue; } // already copied\n\n branch[i].copied = copyNode(branch[i]);\n // now attach the copy to the new 'parellel' tree we are building\n branch[i-1].copied.children.push(branch[i].copied);\n }\n }\n });\n\n depthFirstTraversal(root, function(node, branch) {\n delete node.copied; // cleanup the mutation of the original tree\n });\n\n return filteredResult;\n}\n\nfunction filterTreeList(list, ids) {\n var filteredList = [];\n for(var i = 0, len = list.length; i < len; i++) {\n filteredList.push( filterTree(list[i], ids) );\n }\n return filteredList;\n}\n\nvar hierarchy = [ /* your data here */ ];\nvar ids = [1,3];\n\nvar filtered = filterTreeList(hierarchy, ids);\n\n"
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"Are the midterm results really a victory for female voters?",
"\n\nFollow the author of this article\n\nFollow the topics within this article\n\nAlexandria Ocasio-Cortez, 29, has been elected to the House of Representatives\n\nIt’s a great time to be a woman with political ambitions in the US - as long as you’re a Democrat. ",
"People are rightly talking about the success of the 'pink vote'; the record number of women who have won seats in Congress in the midterm elections, but across the board, there's a less gender balanced picture. ",
"There’s been a fall in the number of Republican women in the House of Representatives, something that reflects the extremely polarised nature of American politics in the era of Donald Trump.",
"\n\nThere’s no doubt that the President’s apparent contempt for women has not just energised millions of female voters, but encouraged many to put themselves forward as candidates. ",
"Yesterday’s elections brought..."
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"Polyembryoma of ovary producing alpha-fetoprotein and HCG: immunoperoxidase and electron microscopic study.",
"\nPolyembryoma of ovary producing alpha-fetoprotein (AFP) and human chorionic gonadotropin (HCG) was studied by indirect immunoperoxidase method for AFP and HCG, and electron microscopy. ",
"Clinically, this patient showed pseudoprecocious puberty caused by elevated HCG which is synthesized by trophoblastic cells in polyembryoma. ",
"She is put under VAC (vincristine, actinomycin-D, cyclophosphamide) chemotherapy after operation and shows no signs of recurrence including reelevation of serum AFP at five months after operation. ",
"Embryoid bodies which we studied correspond to normal embryo at 15- or 16-day stage. ",
"Immunoperoxidase study showed that AFP is synthesized by yolk sac cells of the embryoid bodies and HCG is synthesized by syncytiotrophoblastic cells. ",
"The finding about AFP synthesis suggests that normal embryo at 15- or 16-day stage may begin AFP synthesis. ",
"Electron microscopic study showed that each part of the embryoid bodies had some characteristic structures. ",
"Most striking features found in the cytodifferentiation of the embryoid bodies were noticed in some special differentiation of plasma membrane and existence of surface coat. ",
"Desmosomes were found in endodermal cells and yolk sac cells. ",
"Ectodermal cells were attached to each other by zonulae occludentes and adherentes. ",
"Microvilli were found in ectodermal cells and yolk sac cells. ",
"Two different kinds of surface coat were found in mesodermal cells and lining cells of yolk sac cavity: thin-layered deposit of electron-dense material covering the plasma membrane facing intercellular space of mesodermal cells and endodermal cells, and thick-layered deposit of electron-dense material which covered the plasma membrane facing yolk sac cavity of endodermal cells and yolk sac cells. ",
"Presence of similar characteristic material in RER of yolk sac cells led us to speculate that thick deposit was synthesized by yolk sac cells and secreted into yolk sac cavity. ",
"Combined with immunoperoxidase study by light microscopy, we assume that this thick-layered deposit has some close relation to AFP."
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[
"American coot\n\nThe American coot (Fulica americana), also known as a mud hen or pouldeau, is a bird of the family Rallidae. ",
"Though commonly mistaken for ducks, American coots are only distantly related to ducks, belonging to a separate order. ",
"Unlike the webbed feet of ducks, coots have broad, lobed scales on their lower legs and toes that fold back with each step in order to facilitate walking on dry land. ",
"Coots live near water, typically inhabiting wetlands and open water bodies in North America. ",
"Groups of coots are called covers or rafts. ",
"The oldest known coot lived to be 22 years old.",
"\n\nThe American coot is a migratory bird that occupies most of North America. ",
"It lives in the Pacific and southwestern United States and Mexico year-round and occupies more northeastern regions during the summer breeding season. ",
"In the winter they can be found as far south as Panama. ",
"Coots generally build floating nests and lay 8–12 eggs per clutch. ",
"Females and males have similar appearances, but they can be distinguished during aggressive displays by the larger ruff (head plumage) on the male. ",
"American coots eat primarily algae and other aquatic plants but also animals (both vertebrates and invertebrates) when available.",
"\n\nThe American coot is listed as “Least Concern” under the IUCN conservation ratings. ",
"Hunters generally avoid killing American coots because their meat is not as sought after as that of ducks.",
"\n\nMuch research has been done on the breeding habits of American coots. ",
"Studies have found that mothers will preferentially feed offspring with the brightest plume feathers, a characteristic known as chick ornaments. ",
"American coots are also susceptible to conspecific brood parasitism and have evolved mechanisms to identify which offspring are theirs and which are from parasitic females.",
"\n\nTaxonomy and systematics\n\nThe American coot was first described by Johann Friedrich Gmelin in 1789. ",
"A member of the family Rallidae, it has three subspecies in addition to the nominate: Fulica americana alai, F. a. caribaea, and F. a. ardesiaca. ",
"These subspecies, however, are sometimes considered to be their own separate species. ",
"Including these 'subspecies', there are 11 members of the genus Fulica distributed across the globe.",
"\n\nCoot fossils from the Middle Pleistocene of California have been described as Fulica hesterna but cannot be separated from the present-day American coot. ",
"However, the Pleistocene coot Fulica shufeldti (formerly F. minor), famously known as part of the Fossil Lake fauna, quite possibly was a paleosubspecies of the American coot (as Fulica americana shufeldti) as they only differed marginally in size and proportions from living birds. ",
"Thus, it seems that the modern-type American coots evolved during the mid-late Pleistocene, a few hundred thousand years ago.",
"\n\nThe American coot's genus name, Fulica, is a direct borrowing of the Latin word for \"coot\". ",
"The specific epithet americana means \"America\".",
"\n\nDescription\nThe American coot measures in length and across the wings. ",
"Adults have a short, thick, white bill and white frontal shield, which usually has a reddish-brown spot near the top of the bill between the eyes. ",
"Males and females look alike, but females are smaller. ",
"Body mass in females ranges from and in males from . ",
"Juvenile birds have olive-brown crowns and a gray body. ",
"They become adult-colored around 4 months of age.",
"\n\nVocalizations\nThe American coot has a variety of repeated calls and sounds. ",
"Male and female coots make different types of calls to similar situations. ",
"Male alarm calls are puhlk while female alarm calls are poonk. ",
"Also, stressed males go puhk-cowah or pow-ur while females call cooah.",
"\n\nDistribution and habitat\nAmerican coots are found near water reed-ringed lakes and ponds, open marshes, and sluggish rivers. ",
"They prefer freshwater environments but may temporarily live in saltwater environments during the winter months.",
"\n\nThe American coot's breeding habitat extends from marshes in southern Quebec to the Pacific coast of North America and as far south as northern South America. ",
"Birds from temperate North America east of the Rocky Mountains migrate to the southern United States and southern British Columbia. ",
"It is often a year-round resident where water remains open in winter. ",
"The number of birds that stay year-round near the northern limit of the species' range seems to be increasing.",
"\n\nAutumn migration occurs from August to December, with males and non-breeders moving south before the females and juveniles. ",
"Spring migration to breeding ranges occurs from late February to mid-May, with males and older birds moving North first. ",
"There has been evidence of birds travelling as far north as Greenland and Iceland.",
"\n\nCaribbean coot\n\nCoots resident in the Caribbean and Greater and Lesser Antilles lack the red portion of the frontal shield, and were previously believed to be a distinct species, the Caribbean coot (Fulica caribaea). ",
"In 2016, due to research showing that the only distinguishing characteristic between American and Caribbean coots, the presence or absence of red in the frontal shield, was not distinct to Caribbean coots as some American coots, in locations where vagrancy from Caribbean populations was highly unlikely, had fully white shields and, therefore, there was no way to reliably distinguish the species, and there was no evidence of Caribbean and American coots engaging in assortative mating, the American Ornithological Society lumped the Caribbean coot as a regional variation of the American coot.",
"\n\nBehavior and ecology\nThe American coot is a highly gregarious species, particularly in the winter, when its flocks can number in the thousands. ",
"When swimming on the water surface, American coots exhibit a variety of interesting collective formations, including single-file lines, high density synchronized swimming and rotational dynamics, broad arcing formations, and sequential take-off dynamics.",
"\n\nFeeding\nThe American coot can dive for food but can also forage and scavenge on land. ",
"Their principal source of food is aquatic vegetation, especially algae. ",
"Yet they are omnivorous, also eating arthropods, fish, and other aquatic animals. ",
" During breeding season, coots are more likely to eat aquatic insects and mollusks—which constitute the majority of a chick's diet.",
"\n\nBreeding\n\nThe coot mating season occurs during May and June. ",
"Coot mate pairings are monogamous throughout their life, given they have a suitable territory. ",
"A typical reproductive cycle involves multiple stages: pairing, nesting, copulation, egg deposition, incubation, and hatching.",
"\nThe American coot typically has long courtship periods. ",
"This courtship period is characterized by billing, bowing, and nibbling. ",
"Males generally initiate billing, which is the touching of bills between individuals. ",
"As the pair bond becomes more evident, both males and females will initiate billing only with each other and not other males or females. ",
"After a pair bond is cemented, the mating pair looks for a territory to build a nest in. ",
"A pair bond becomes permanent when a nesting territory is secured. ",
"Copulation behavior among coot pairs always falls under the same general pattern. ",
"First the male chases the female. ",
"Then, the female moves to the display platform and squats with her head under the water. ",
"The male then mounts the female, using his claws and wings to balance on the female's back while the she brings her head above the water. ",
"Sexual intercourse usually takes no longer than two seconds.",
"\n\nNests\nThe American coot is a prolific builder and will create multiple structures during a single breeding season. ",
"It nests in well-concealed locations in tall reeds. ",
"There are three general types of structures: display platforms, egg nests and brood nests.",
"\n Display platforms are used as roosting sites and are left to decompose after copulation. ",
"\n Egg nests are typically in diameter with a ramp that allows the parents to enter and exit without tearing the sides of the nests. ",
"Coots will often build multiple egg nests before selecting one to lay their eggs in. ",
"\n Brood nests are nests that are either newly constructed or have been converted from old egg nests after the eggs hatch, becoming larger egg nests. ",
"\nSince American coots build on the water, their structures disintegrate easily and have short life spans. ",
"Egg and brood nests are actually elaborate rafts, and must be constantly added to in order to stay afloat. ",
"Females typically do the most work while building.",
"\n\nEgg-laying and clutch size\nFemales deposit one egg a day until the clutch is complete. ",
"Eggs are usually deposited between sunset and midnight. ",
"Typically, early season and first clutches average two more eggs than second nestings and late season clutches. ",
"Early season nests see an average of 9.0 eggs per clutch while late clutches see an average of 6.4 eggs per clutch. ",
"There is an inverse relationship between egg weights and laying sequence, wherein earlier eggs are larger than eggs laid later in the sequence. ",
"It is possible to induce a female coot to lay more eggs than normal by either removing all or part of her clutch. ",
"Sometimes, a female may abandon the clutch if enough eggs are removed. ",
"Coots, however, do not respond to experimental addition of eggs by laying fewer eggs.",
"\n\nThe American coot is a persistent re-nester, and will replace lost clutches with new ones within two days of clutch-loss during deposition. ",
"One study showed that 68% of destroyed clutches are eventually replaced. ",
"Re-nested clutches are typically smaller than original clutches by one or two eggs, but this could be attributed to differences in time and habitat quality instead of food or nutrient reserves and availability.",
"\n\nYounger females reproduce later in the season and produce smaller eggs than older females. ",
"Their offspring are also smaller. ",
"However, there is no difference in clutch size between older and younger females as there is in other avian species.",
"\n\nIncubation and hatching\nIncubation start time in the American coot is variable, and can begin anywhere from the deposition of the first egg to after the clutch is fully deposited. ",
"Starting incubation before the entire clutch has been laid is an uncommon practice among birds. ",
"Once incubation starts it continues without interruption. ",
"Male and female coots share incubation responsibility, but males do most of the work during the 21-day incubation period. ",
"Females will begin to re-nest clutches in an average of six days if clutches are destroyed during incubation.",
"\n\nHatch order usually follows the same sequence as laying order. ",
"Regardless of clutch size, eight is the typical maximum size of a brood. ",
"Egg desertion is a frequent occurrence among coots because females will often deposit more than eight eggs. ",
"Brood size limits incubation time, and when a certain number of chicks have hatched the remaining eggs are abandoned. ",
"The mechanism for egg abandonment has not yet been discovered. ",
"Food resource constraints may limit the number of eggs parents let hatch, or the remaining eggs may not provide enough visual or tactile stimulation to elicit incubation behavior. ",
"An American coot can be forced to hatch more eggs than are normally laid. ",
"These additional offspring, however, suffer higher mortality rates due to inadequacy in brooding or feeding ability.",
"\n\nMaternal effects\nHormones that are passed down from the mother into the egg affect offspring growth, behavior, and social interactions. ",
"These nongenetic contributions by the mother are known as maternal effects. ",
"In the American coot, two levels of androgen and testosterone variation have been discovered—within-clutch and among-clutch variation. ",
"Within the same clutch, eggs laid earlier in the sequence have higher testosterone levels than eggs laid later in the sequence. ",
"Females that lay three clutches deposit more androgens into their yolks than females who lay only one or two clutches.",
"\n\nBrood parasitism\nThe American coot has a mixed reproductive strategy, and conspecific brood parasitism is a common alternative reproductive method. ",
"In one 4-year study, researchers found that 40% of nests were parasitized, and that 13% of all eggs were laid by females in nests that were not their own. ",
"Increasing reproductive success under social and ecological constraints is the primary reason for brood parasitism. ",
"Floater females without territories or nests use brood parasitism as their primary method of reproduction, if they breed at all. ",
"Other females may engage in brood parasitism if their partially complete clutches are destroyed. ",
"Conspecific brood parasitic behavior is most common among females trying to increase their total number of offspring. ",
"Food supply is the limiting factor to chick survival and starvation is the most common cause of chick morbidity. ",
"Parasitic females bypass the parental care constraint of feeding by laying additional parasitic eggs in addition to their normal nest.",
"\n\nWhen a parasitic female lays her egg in a host female's nest, the host female experiences a deposition rate of two eggs per day. ",
"Host females may recognize parasitic eggs when the egg deposition pattern deviates from the traditional one egg per day pattern. ",
"The occurrence of brood parasitism may be influenced by the body size of the potential parasitic female relative to the potential host female. ",
"Parasitic females are generally larger than their host counterparts, but on average, there is no size difference between the parasite and the host.",
"\n\nThe American coot, unlike other parasitized species, has the ability to recognize and reject conspecific parasitic chicks from their brood. ",
"Parents aggressively reject parasite chicks by pecking them vigorously, drowning them, preventing them from entering the nest, etc. ",
"They learn to recognize their own chicks by imprinting on cues from the first chick that hatches. ",
"The first-hatched chick is a reference to which parents discriminate between later-hatched chicks. ",
"Chicks that do not match the imprinted cues are then recognized as parasite chicks and are rejected.",
"\n\nChick recognition reduces the costs associated with parasitism, and coots are one of only three bird species in which this behavior has evolved. ",
"This is because hatching order is predictable in parasitized coots—host eggs will reliably hatch before parasite eggs. ",
"In other species where hatching order is not as reliable, there is a risk of misimprinting on a parasite chick first and then rejecting their own chicks. ",
"In these species, the cost of accidentally misimprinting is greater than the benefits of rejecting parasite chicks.",
"\n\nChick ornaments\n\nThe first evidence for parental selection of exaggerated, ornamental traits in offspring was found in American coots. ",
"Black American coot chicks have conspicuously orange-tipped ornamental plumes covering the front half of their body that are known as “chick ornaments” that eventually get bleached out after six days. ",
"This brightly colored, exaggerated trait makes coot chicks more susceptible to predation and does not aid in thermoregulation, but remains selected for by parental choice. ",
"These plumes are not necessary for chick viability, but increased chick ornamentation increases the likelihood that a chick will be chosen as a favorite by the parents. ",
"Experimental manipulation of chick ornamentation by clipping the bright plumes has shown that parents prefer ornamented chicks over non-ornamented ones.",
"\n\nPredation\nThe American coot is fairly aggressive in defense of its eggs and, in combination with their protected nesting habitat, undoubtedly helps reduce losses of eggs and young to all but the most determined and effective predators. ",
"American crows, black-billed magpies and Forster's tern can sometime take eggs. ",
"Mammalian predators (including red foxes, coyotes, skunks and raccoons) are even less likely to predate coot nests, though nests are regularly destroyed in usurpation by muskrats. ",
"Conversely, the bold behavior of immature and adult coots leads to them falling prey with relative regularity once out of the breeding season. ",
"Regular, non-nesting-season predators include great horned owls, northern harriers, bald eagles, golden eagles, American alligators, bobcats, great black-backed and California gulls. ",
"In fact, coots may locally comprise more than 80% of the bald eagle diet.",
"\n\nIn culture\nOn the Louisiana coast, the Cajun word for coot is pouldeau, from French for \"coot\", poule d'eau – literally \"water hen\". ",
"Coot can be used for cooking; it is somewhat popular in Cajun cuisine, for instance as an ingredient for gumbos cooked at home by duck hunters.",
"\n\nThe bird is the mascot of the Toledo Mud Hens Minor League Baseball team.",
"\n\nConservation and threats\nThe American coot is listed under \"least concern\" by the IUCN Red List of Endangered Species. ",
"They are common and widespread, and are sometimes even considered a pest. ",
"They are rarely the targets of hunters since their meat is not considered to be as good as that of ducks; although some are shot for sport, particularly in the southeastern United States. ",
"Because they are found in wetlands, scientists use them to monitor toxin levels and pollution problems in these environments.",
"\n\nReferences\n\nCited sources\n\nExternal links\n\n American Coot – Fulica americana – USGS Patuxent Bird Identification InfoCenter.",
"\n American Coot Species Account – Cornell Lab of Ornithology\n \n \n\nAmerican coot\nAmerican Coot\nCategory:Birds of North America\nCategory:Birds of the United States\nCategory:Birds of Central America\nAmerican coot"
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"Chelsea FC\nTickets\n\nFounded in 1905, Chelsea FC continues to hold ground as one of the most famous and respected club football teams in the entire world, and it’s certainly for good reason. ",
"The Blues have claimed multiple English Premier League titles, and they’ve left their mark throughout European soil as well. ",
"London’s finest is no stranger to claiming UEFA Champions League titles, and the roar created every time the team steps inside Stamford Bridge epitomizes the global passion for “The Beautiful Game.” ",
"Throughout its history, Chelsea football has featured many of the game’s top players, including Peter Osgood, Gianfranco Zola, Didier Drogba, Frank Lampard, and John Terry, and a purchase of Chelsea tickets will allow you to witness this iconic squad in action.",
"\n\nChelsea FC Soccer Tickets from VividSeats.com\n\nDiscover Vivid Seats’ online catalog of available Chelsea soccer tickets today, and set aside seats for the Blues upcoming English Premier League campaign. ",
"Support Chelsea at home at Stamford Bridge and on the road this year! ",
"Winning three EPL titles in the 2000’s, Chelsea has become one of the strongest clubs in Europe. ",
"The club’s massive fan base around the world has caused the price of tickets for Chelsea soccer to rise dramatically. ",
"Vivid Seats, nonetheless, can offer you reasonably priced seats at the Chelsea soccer game of your choosing for EPL, FA Cup, and Champions League matches.",
"\n\nVivid Seats customers enjoy the benefit of having our dedicated team of service representatives always available to answer any questions. ",
"You can engage a live agent on VividSeats.com about purchasing Chelsea FC tickets in seconds. ",
"Simply select the icon labelled ‘Live Chat’ at the top of the screen, and converse with a representative from either our sales, technical support, or customer service departments.",
"\n\nBook Chelsea soccer tickets today to see top-flight football at the world famous Stamford Bridge. ",
"Stamford Bridge, a UEFA Elite Stadium, seats over 41,000 supporters on match day. ",
"Watch the Blues go up against top EPL sides like Manchester United, Manchester City, Arsenal, Tottenham, and Liverpool at “The Bridge” by purchasing Chelsea soccer tickets from Vivid Seats.",
"\n\nSoccer\nNews\n\nChelsea vs. Manchester City Coming to MLB Stadiums in May 2013\n\nFans of English Premier League football will get their chance to witness two of the league’s finest clubs live in the U.S. this spring, as Chelsea FC will face Manchester City FC at Busch Stadium in St. Louis, Mo., on May 23, followed by a match at Yankee Stadium in New York City on May 25. ",
"The Busch Stadium game will mark a milestone for the venue, as it will be the first professional sport other than baseball to take place inside the home of Major League Baseball’s St. Louis Cardinals, while the Yankee Stadium contest will be the second consecutive summer that Chelsea plays at the home of the New York Yankees. ",
"In July 2012, Chelsea tied Paris Saint-Germain, 1-1, in front of over 38,000 fans. ",
"The Blues stand as the defending UEFA Champions League winners, thanks to the golden foot of Didier Drogba, while Manchester City is the current Premier League champion, so fans can expect two fully packed stadiums when these legendary clubs face off.",
"\n\nSpectators will witness a bevy of the best players in the world showing off their skills in this highly anticipated friendly, as both teams feature more top-flight talent between them than many leagues boast in total. ",
"Fans of Chelsea football are surely pumped to get a first-hand glimpse at stars like Juan Mata, Frank Lampard, Ashley Cole, Oscar, Eden Hazard, and Ramires, while Manchester City fans can look forward to superstars like Yaya Toure, Samir Nasri, Carlos Tevez, Sergio Kun Aguero, Joe Hart, and Vincent Kompany. ",
"Those looking to purchase Chelsea tickets can also look forward to the team’s incredible matchups in the English Premier League, as the Blues will be taking on squads like Liverpool FC on April 17, Manchester United on May 4, and U.S. Men’s National Team goalie Tim Howard’s club, Everton, on May 19."
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"Ferlacher Badesee\n\nFerlacher Badesee is a lake of Carinthia, Austria.",
"\n\nCategory:Lakes of Carinthia (state)"
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"[Comparative estimation of traumaticity of laparoscopic and open cholecystectomy according to objective methods of investigation].",
"\nThe results of clinic-laboratory investigations were adduced, which confirm objectively the lesser traumaticity of laparoscopic cholecystectomy (LCE) in comparison with open cholecystectomy in patients, suffering an acute cholecystitis. ",
"While investigation of the patients blood serum, in whom LCE or open cholecystectomy was performed, using laser correlation spectroscopy (LCS), and performance of malonic dialdehide and dienic conjugate content determination, more rapid restoration of homeostasis parameters was proved after LCE, trusting its lesser traumaticity."
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[
"Introduction\n============\n\nAltered pericellular protease activity is a critical component of the metastatic cascade in many tumour types \\[[@b1]-[@b5]\\]. ",
"The protease activity that accompanies tumour invasion promotes cell migration by digesting physical barriers, exposing cryptic binding sites, liberating embedded growth/regulatory factors and simultaneously decreasing the availability of matrix adhesion ligands \\[[@b6]-[@b8]\\]. ",
"Cellular invasion correlates with matrix metalloproteinase-1 (MMP-1) expression in chondrosarcoma cells and transient down-regulation of MMP-1 expression decreases the cell invasion *in vitro* \\[[@b1], [@b9], [@b10]\\]. ",
"Despite the wealth of pre-clinical data implicating MMP-1 as a therapeutic target, the clinical trials with MMP inhibitors in cancer therapy provided disappointing results \\[[@b11]-[@b13]\\]. ",
"The reasons for this maybe several fold but likely include an attempt to indiscriminately inhibit a process that is not completely understood; namely, the regulation of MMP intra- and extracellular activity, production, delivery, compartmentalization and activation of this group of proteases \\[[@b14]\\].",
"\n\nInvestigators have examined cancer cell migration and pericellular proteolysis with sophisticated imaging techniques \\[[@b15],[@b16]\\]. ",
"They have demonstrated that MMPs are secreted in very specific pericellular locations and that these had biological and mechanical consequences for directed cell movement \\[[@b17]-[@b20]\\]. ",
"These studies support the contention that indiscriminate inhibition of MMPs determined the unsuccessful fate of previous clinical trials \\[[@b11]-[@b13]\\]. ",
"This interpretation led our laboratory and others to postulate that a more complete understanding of post-translational modification and delivery of MMPs would permit the development of a successful clinical strategy for novel MMP inhibitors \\[[@b14]\\].",
"\n\nPrenylation facilitates protein attachment to cell membrane \\[[@b21]\\]. ",
"It involves a 15-carbon farnesyl (FT) or 20-carbon geranylgeranyl (GGT) isoprenoid tag attachment to the target protein carboxyl-terminal cysteine residues on preferred CAAX target sequences. ",
"This process is catalysed by enzyme complexes termed protein farnesyltransferase (FTase) and protein geranylgeranyltransferase type I and II (GGTase-I and II) \\[[@b22]-[@b25]\\]. ",
"Inhibition of prenylation has been explored as an anti-neoplastic strategy in various cancers, affecting numerous cellular processes and signalling cascades including Ras \\[[@b26]-[@b29]\\]. ",
"Prenylation inhibitors have also been reported to disrupt subcellular trafficking of proteins within cells \\[[@b30]\\]. ",
"These interventions reduced tumour burden and induced apoptosis *in vitro* and in pre-clinical models \\[[@b31],[@b32]\\]. ",
"The specific mechanisms for the observed anti-neoplastic effects were unclear because of the breadth of protein targets of prenylation \\[[@b33]-[@b37]\\]. ",
"Recently investigators have reported that in rheumatoid arthritis, MMP-1 secretion from synovial tissue could be inhibited by blocking prenylation \\[[@b38]\\]. ",
"The study did not specifically investigate the effect of inhibition on MMP-1 subcellular delivery, documenting only affects on general secretion. ",
"It should be noted that MMP-1 by itself is not prenylated (there is no existence of suitable carboxyl-terminal target sequence). ",
"Based on what is known about prenylation and protein trafficking, inhibition of MMP-1 directional traffic is likely to have important effects on cell migration and tumour invasion particularly in human chondrosarcoma \\[[@b39]-[@b42]\\].",
"\n\nThis study demonstrates that the ability of a cell to invade a collagen barrier is partially related to MMP-1 delivery to podia structures. ",
"Inhibition of prenylation affects lamellipodia formation, MMP-1 localization into these structures and secretion. ",
"The lamellipodia formation can be partially restored by the prenylation agonist farnesyl pyrophosphate (FPP), while MMP-1 delivery to these structures delayed under the time frame investigated. ",
"This study seeks to understand the intracellular directional delivery of MMP-1 in support of a better devised and targeted approach to MMP inhibition.",
"\n\nMaterials and methods\n=====================\n\nCell culture and inhibition of prenylation\n------------------------------------------\n\nHuman osteogenic sarcoma cells (143B, CRL-8303; ATCC Bethesda, MD, USA) were cultured in DMEM (\\#10-017CV; Invitrogen, Carlsbad, CA, USA) complemented with 10% foetal bovine serum (FBS). ",
"After trypsinization, cells were quenched in DMEM with 5% bovine serum albumin then incubated overnight in p100 plates at 37°C to achieve a 70% confluence before serum starvation. ",
"Next FTI-276 {15 μM, \\[4-\\[2-(*R*)-amino-3-mercaptopropyl\\]amino-2-phenylbenzoyl\\] methionine trifluoroacetate salt, \\#F9553; Sigma-Aldrich, St. Louis, MO, USA} and/or its antagonists FPP (20 μM, 3,7,11-Trimethyl-2,6,10-dodecatrien-1-yl pyrophosphate ammonium salt, \\#F6892; Sigma-Aldrich) alone, or in combination of two were used in serum-free DMEM overnight. ",
"Following exposure, cells were washed with PBS and harvested on ice. ",
"Samples either were processed immediately or snap frozen in liquid N~2~ and stored at −80°C until further use.",
"\n\nGeneration of stable RNAi clones\n--------------------------------\n\nStable clones were generated by pHuSH 29mer shRNA constructs targeting MMP-1 (\\#TR311450; OriGene, Rockville, MD, USA) and established by puromycin selection (Invitrogen; 1.0 μg/ml in the media above) on six-well plates. ",
"The RNAi kit composed of three separated targeting sequences and a negative control. ",
"Individual colonies were isolated through parallel process, expanded in selection media containing the antibiotics until passage was transferred into p100 plates. ",
"Then the antibiotic was removed, and colonies were screened for gene expression changes by RT-qPCR. ",
"The screening process was repeated at least three to five times with every second passages to establish the stability of down-regulation. ",
"Clones used in this study were designated as T6-4, T6-5 and T6-7.",
"\n\nCollagen-based cell invasion assays\n-----------------------------------\n\nQCM™ 96-well collagen-based cell invasion assay from Chemicon International (\\#ECM556) was used. ",
"Coatings on the plates were avian collagens Type I (85%) and Type III (15%), respectively. ",
"Assays were performed according to the protocol's manual with the following modifications: after initial cell number normalization, 4.0 χ10^4^ cells/well was seeded onto the plates (nine parallels with three blank controls) in media combined with pharmacologic agents (untreated controls, FTI-276, FPP, FTI-276/FPP). ",
"Feeder wells contained the same media ± 10% FBS to determine the effects of natural metalloproteinase inhibitors and incubated overnight. ",
"The following day the plates were processed according to the Chemicon's protocol then measured with 480/520 nm filter set on Berthold Mithras LB 970 plate reader. ",
"The data were transferred into Microsoft Excel for further statistical analysis (ANOVA; where the null hypothesis is that conditions are different and confirmed when *P* \\< 0.05) and graphing.",
"\n\nThree-dimensional collagen gel matrices and invadopodia assay\n-------------------------------------------------------------\n\nOne packet of alpha Minimal Essential Medium (α-MEM, \\#11900-073; Invitrogen) was dissolved in 100 ml of endotoxin-free, distilled water to bring the medium concentration to 10-fold. ",
"2.2 g of sodium bicarbonate was added to neutralize the media. ",
"Purcol acidified collagen (97% Type I collagen/3% Type III collagen, 3.0 mg/ml, \\#5409; INAMED Corporation, Santa Barbara CA, USA) was aliquoted into 15 ml conical tubes for individual use. ",
"Subsequently, collagen gels were mixed in the following proportions: 1000 μl Purcol Collagen, 125 μl α-MEM and 175 μl DQ Collagen (\\#D12060; Invitrogen). ",
"The solution was neutralized to pH 7.0 by the addition of 0.8M sodium bicarbonate prepared in endotoxin-free distilled water. ",
"Gels were cross-linked with 0.3% glutaraldehyde/PBS to the cover slips (5 min. ",
"at room temperature and 15 min. ",
"on ice). ",
"Total gel volume of 70 μl was used with 12-mm circular cover slips (polymerized on parafilm upside down) to standardize its thickness to ∼80--100 μm. ",
"Gels were then seeded at 2.5 χ 10^4^ cells/ml/well concentration. ",
"For lamellipodia MMP-1--specific activity studies, Fluorogenic MMP1-Substrate-III (\\#444219; Calbiochem, EMD Biosciences, Gibbstown NJ, USA) was used (1 mg/ml) to saturate the gels. ",
"The MatTek dishes (P35G-1.0-14-C) were prepared as described \\[[@b43]-[@b45]\\] with 150 μl of matrix mixture. ",
"Excess was aspirated and dishes dried in dark for 30 min. ",
"followed by cross-linking with 2 ml of 0.5% glutaraldehyde/PBS (chilled in ice) and incubated on ice for 15 min., ",
"then at room temperature for 30 min. ",
"Second coating with MMP-1 cleavage specific Fluorogenic Substrate-III was used to saturate the cross-linked gels. ",
"The equilibrated dishes (OptiMEM media, 1% FBS serum) were stored at 4°C in dark until use. ",
"Cell growth, MMP-1 breakdown of DQ-collagen, and MMP1-S-III substrate degradation were observed with Zeiss Axioplan II fluorescent microscope with 40χ objective with DAPI (blue), GFP (green) and DsRed (orange) filters combined with PlasDIC filter (transmitted light and merge) to collect live cell images.",
"\n\nWestern blotting, image acquisition and statistical analysis\n------------------------------------------------------------\n\nExtracellular media was separated from cells by centrifugation and retain for determination of MMP-1. ",
"Cells were then lysed in buffer containing 50 mM Tris--HCl pH 7.4, 1% NP-40, 0.25% Na-deoxycholate, 150 mM NaCl, 1 mM EDTA, 1 mM Na-vanadate and 1 mM phenylmethylsulfonyl fluoride, completed with protease inhibitor cocktail from Roche (Indianapolis, IN, USA). ",
"Samples were normalized for protein content with ND-1000 (NanoDrop Technologies Inc., Wilmington, DE, USA). ",
"For Western blotting, 125 μg protein samples per lane were analysed with primary antibodies \\[MMP-1 \\#sc-58377, farnesyltransferase, CAAX box, alpha (FNTA) \\#sc-136, actin \\#sc-1615; Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA), and \\#610002; BD Biosciences (San Jose, CA, USA)\\], incubated overnight at 4°C, followed by secondary HRP conjugated antibodies (Amersham/GE, Piscataway, NJ, USA) for 1 hr at room temperature. ",
"Bands were detected with SuperSignal West Pico ECL detection kit (Pierce, Rockford, IL, USA) on UVP Biospectrum Digital Imaging System (UVP Inc., Upland, CA, USA). ",
"Raw images were quantized by optical density through supplied densitometry analysis software. ",
"Data were transferred to Excel file and statistical analysis was performed by the ANOVA subroutine of Microsoft Excel and/or MATLAB 7.5.0 software (The MathWorks, Inc., Natick, MA, USA) to verify the significance of densitometry results (0 hypothesis that they are different when *P* \\< 0.05).",
"\n\nReal-time quantitative PCR\n--------------------------\n\nTotal RNAs were purified with RNeasy Mini Kit with on column DNase treatment (Qiagen Inc., Valencia, CA, USA) according to the manufacturer's protocol. ",
"For the cDNA synthesis, 5 μg total RNA was used with High Capacity cDNA Reverse Transcription Kit (\\#4368813; Applied Biosystems, Foster City, CA, USA) as described by the kit manual. ",
"All genes were analysed in triplicate in 20 μl mixture/well containing 100 ng cDNA and 2 TaqMan Fast Universal PCR Master Mix (\\#4352042; Applied Biosystems). ",
"18S was used as the internal control. ",
"Relative RT-qPCR was performed on AB 7900HT Fast Real-Time System using specific human TaqMan gene expression assays MMP-1 (\\#Hs00233958_m1), TIMP1 (\\#Hs00171558_m1), KRAS (\\#Hs00364282_m1), FNTA (\\#Hs00357739_m1). ",
"Relative quantities of each gene were determined by the manufacturer's default ΔΔCt method.",
"\n\nFluorescent microscopy and MetaMorph analysis\n---------------------------------------------\n\nCells were plated in four chamber slides (Lab-Tek 1177399, Nalgene-Nunc Int., ",
"Rochester, NY, USA) at density of (2.5--3.0) χ 10^4^ cells/chamber and then were grown to confluence overnight. ",
"Following exposure to prenylation inhibitors, cells were washed with PBS then fixed by 4% formaldehyde and processed according to standard immunostaining protocols. ",
"Cells were stained with primary antibody against MMP-1 (\\#sc-21731; Santa Cruz), counterstained with DAPI (\\#D21490) and AL647-Phalloidin (\\#A-22287) both from Invitrogen (Molecular Probes, Eugene, OR, USA) for nuclei and F-actin respectively. ",
"The secondary antibody against MMP-1 was AL594 (\\#A-21201) from Molecular Probes. ",
"Slides were mounted in ProLong (\\#P-7481; Molecular Probes) then imaged with Zeiss LSM 510 confocal microscope at Sylvester Comprehensive Cancer Center Bioimaging Core. ",
"Above from the blue, red and far-red (pseudo-coloured as white) channels, reflected interference images from the cover-slips were collected to aid in the determination of lamellipodia regions of the migrating cells. ",
"Confocal z-sections were combined and the resulted images were depth-coded to visualize better the lamellar regions. ",
"Red channel was extracted from the images then total and partial red pixel counts were performed to determine the total MMP-1 levels as well as its localization to lamellipodia structures. ",
"Interference image of same cells with pseudo-coloured intensity levels of MMP-1 (red channel) were depth-coded and used to determine the boundaries of lamellipodia structures. ",
"These regions were marked and numbered in each cell for subsequent pixel counts.",
"\n\nFor MetaMorph analysis, the image background was subtracted in the software by threshold binary processing. ",
"Then lamellipodia regions were outlined (according to the previously depth-coded image), and numbered in the software. ",
"Data were logged by total cell count and region measurement command (lamellipodia pixel count) into Microsoft Excel spreadsheet. ",
"MMP-1 lamellipodia localization and surface area were then related to total cellular MMP-1 and total cell surface and expressed as its percentage. ",
"The counts were performed on 30--50 cells from each condition (untreated, FTI, FPP and FTI/FPP). ",
"For lamellipodia studies, MMP1-Substrate III (excitation/emission ∇ λ 365/450 nm) was recorded with live cells at ∼461 nm (DAPI channel) with Zeiss Axioplan II microscope (40χ objective). ",
"MMP-1 cleavage was quantized in the podia structures of 45--50 cells for each condition through the blue channel.",
"\n\nResults\n=======\n\nCellular invasiveness is affected by farnesyl transferase inhibitor (FTI)\n-------------------------------------------------------------------------\n\nTo investigate how cellular transport of MMP-1 affected tumour cell invasion *in vitro*, we used a prenylation inhibitor (FTI-276) which inactivates the FTase (composed of A and B subunits termed FNTA/FNTB) complex by blocking the active site of the FTase enzyme \\[[@b46]\\]. ",
"To examine the efficacy and specificity of the FT inhibition, cytoplasmic Ras was used as an MMP-1-independent surrogate marker ([Fig. ",
"1A](#fig01){ref-type=\"fig\"}, second row). ",
"FTI-276 prevents Ras farnesylation, efficiently blocking its plasma membrane translocation and consequentially increasing cytoplasmic concentration. ",
"Effects of FTI-276 on both Ras cytoplasmic and extracellular MMP-1 levels are noted between 5 and 15 μM, correlating with inhibition of FTase activity up to 50 μM, indicating that the overall secretion process is prenylation dependent ([Fig. ",
"1A](#fig01){ref-type=\"fig\"}). ",
"Corresponding transmembrane cellular invasion is decreased 25% by FTI-276 consistent with the inhibition of secretion. ",
"FPP (FNTA substrate and prenylation agonist) is able to overcome FNTA inhibition and restore the invasion up to ∼85% to its original levels, indicating that reversal of FT inhibition is associated with partially restored cell function ([Fig. ",
"1B](#fig01){ref-type=\"fig\"}). ",
"Effects of FTI-276 and FPP were significantly different (ANOVA, *P* ∇ 0.001), and null hypothesis confirmed *P-*value \\>0.05 would indicate no difference.",
"\n\n{#fig01}\n\nFarnesyl transferase inhibition decreases MMP-1 extracellular protein levels by 35% and this effect is reversed by FPP to 82%\n\n([Fig. ",
"1C](#fig01){ref-type=\"fig\"}). ",
"FPP alone causes a 20% increase in MMP-1 extracellular protein and leave cytoplasmic Ras levels unchanged ([Fig. ",
"1D](#fig01){ref-type=\"fig\"}, relevant Western blots quantization, *Y*-axis in optical density). ",
"Notably, the addition of FPP- to FTI-276--inhibited cells did not reverse cytoplasmic Ras accumulation and MMP-1 levels were less than FPP alone. ",
"These changes were not associated with significant changes in the levels of FNTA or β-actin.",
"\n\nMMP-1 RNAi validation of FTI affect on cellular invasion\n--------------------------------------------------------\n\nThe question arose as to whether the observed alterations in the extracellular MMP-1 protein levels induced by prenylation inhibition are reflected merely protein export or whether there is an associated change in gene expression too ([Fig. ",
"2A](#fig02){ref-type=\"fig\"}). ",
"FTI-276 down-regulated MMP-1 gene expression by 29.84% and up-regulated Ras mRNA levels by 23.42% (used only as indicator of the inhibitor effectiveness at mRNA level). ",
"TIMP1 (a natural MMP-1 inhibitor) is demonstrates parallel changes with MMP-1 gene expression. ",
"FNTA expression was not affected significantly by prenylation inhibition or substrate excess. ",
"Differential magnitude of changes between gene expression and secretion indicate that they might be guided by different kinetics. ",
"To validate further, the MMP-1 contribution to cellular invasiveness OriGene pHuSH targeting MMP-1 constructs were used to establish three parallel stable cell lines (143B cells; [Fig. ",
"2B](#fig02){ref-type=\"fig\"}). ",
"Following the selection process, clones showing over 80% stable MMP-1 down-regulation were designated as T6-4, T6-5 and T6-7. ",
"Targeting specificity was confirmed through MMP-14 (MT-MMP1), which importantly also associated with the invasion process. ",
"It was slightly up-regulated in two clones and down-regulated (∼30%) in T6-7. ",
"This slight up-regulation did not rescue the MMP-1 knockdown effect on the cellular invasiveness ([Fig. ",
"2D](#fig02){ref-type=\"fig\"}). ",
"MMP-1 protein expression levels confirm the RT-qPCR analysis ([Fig. ",
"2C](#fig02){ref-type=\"fig\"}). ",
"Here the equal loading was confirmed by β-actin. ",
"Over 80% of MMP-1 mRNA down-regulation at the gene expressional level in all three clones ([Fig. ",
"2B](#fig02){ref-type=\"fig\"}) was responsible for ∼50% decrease in trans-membrane invasion/migratory capacity ([Fig. ",
"2D](#fig02){ref-type=\"fig\"}; ANOVA**,***P* \\< 0.001; T6-4 and T6-7 invasion levels statistically similar, *P* ∇ 0.129).",
"\n\n{#fig02}\n\nMMP-1 cellular lamellipodia distribution is FTI sensitive\n---------------------------------------------------------\n\nCellular transmembrane invasion reflects and measures invadopodia-related MMP activity and matrix degradation. ",
"The observed MMP-1 contribution to *in situ* invasion results between RNAi interference and FTI-276 treatment ([Figs 1](#fig01){ref-type=\"fig\"} and [2](#fig02){ref-type=\"fig\"}) raised the possibility that sub-cellular distribution of MMP-1 might be an important component of the invasive mechanism(s). ",
"To further understand the relationship between MMP-1 localization and invasion, we examined how FNTA inhibition affects cell morphology. ",
"First, to determine the extent of the lamellipodia for each cell ([Fig. ",
"3A](#fig03){ref-type=\"fig\"}), the cells were evaluated with fluorescence microscopy. ",
"Image analysis was aided by DAPI staining to locate the cell nucleus and Alexa-647-Phalloidin staining of the stress fibres. ",
"Depth-coding visualized the lamellipodia area as relatively smooth border regions in comparison to the rest of the cell body. ",
"The boundaries were then traced (G) and areas numbered and quantized with MetaMorph software. ",
"Quantization of lamellipodia surface areas and containing MMP-1 pixels ([Fig. ",
"3B](#fig03){ref-type=\"fig\"}) show that FPP increases both lamellipodia formation and MMP-1 localization ([Fig. ",
"3B](#fig03){ref-type=\"fig\"}, ∼24% and ∼19%, respectively). ",
"The surface area of lamellipodia changed with FTI-276, as did the MMP-1 localization within lamellipodia ([Fig. ",
"3B](#fig03){ref-type=\"fig\"}). ",
"Results of combined FPP/FTI exposure indicate that while lamellipodia formation was partially restored (87%), the lamellipodia MMP-1 localization was not under the investigated time period. ",
"It was observed that FTI-276 results in depletion of MMP-1 in the lamellipodia area by 60% when compared to untreated cells ([Fig. ",
"3B](#fig03){ref-type=\"fig\"}). ",
"FPP was not able to rescue the delivery of MMP-1 to the cell periphery. ",
"The histograms demonstrate significant differences between pharmacologic agents for lamellipodia formation and for MMP-1 content (ANOVA, *P* \\< 0.0001). ",
"This result is in agreement with observed changes in extracellular MMP-1 protein levels and invasive behaviour of the sarcoma cells.",
"\n\n![",
"Determination of MMP-1 lamellipodia localization. (",
"A) Visual demonstration of objective invadopodia boundary determination through depth-coding. (",
"A) Cellular architecture was outlined by Alexa-647-phalloidin staining (pseudo-coloured white) and aided by nuclear counterstaining with DAPI (B-blue). ",
"MMP-1 localization (C-red) and the reflected image (D-turquoise). ",
"The reflection image interference rings \\[E-Merge, labelled with asterisk (~\\*~)\\] delineate the stretched out regions of cellular bodies (lamellipodia) showing striped wave-like formations of protruding cellular frontlines. ",
"To demonstrate this phenomena more clearly, the interferential image was pseudo-coloured according to the height of cellular features (depth-coding, F). ",
"The boundaries of lamellipodia region circled (G-white). (",
"B) MMP-1 delivery is FTI-276 sensitive. ",
"Fifteen to twenty randomly chosen fields of individual images (representing 60--90 individual cells) were captured of each condition with 63χ oil immersion lens on the Zeiss LSM510 confocal microscope. ",
"Blue (DAPI, nucleus), red (MMP-1), pseudo-coloured white (F-actin) and reflected interference images (not shown) were collected. ",
"MetaMorph analysis was performed on the extracted red channels for total and partial surface areas and pixel counts to determine the podia formation activity and its partial MMP-1 representation as described. ",
"Each cell total surface area and MMP-1 content treated as fixed (100%). ",
"From this the lamellipodia formation and partial MMP-1 localization were determined as percentage of total values (surface area and red pixel counts). ",
"These percentage results presented as bar graph, where untreated cells provide the reference basis ∇ 100% showing for drugs affecting lamellipodia surface area (bar graph on the left) and MMP-1 loading (bar graph on the right). ",
"Statistical analysis shows that the quantization of lamellipodia area and MMP-1 loading are different for and by the presented conditions with *P* ∇ 4.09E−73 and *P* ∇ 5.60786E−30 confidence levels, respectively.](jcmm0016-1331-f3){#fig03}\n\nInvadopodia MMP-1 activity levels are decreased by FTI\n------------------------------------------------------\n\nGeneral paradigm is that MMP-1 secreted and deposited in the extracellular matrix as an inactive zymogen by various cell types where it is subsequently activated \\[[@b47]\\]. ",
"To investigate further if MMP-1 cellular production and localization can overlap with the underlying collagen matrix degradation beneath the cells, live cell fluorescent imaging in combination with DQ-collagen, MMP1-DsRed fusion protein expression and MMP-1 cleavage specific Fluorogenic Substrate-III ([Fig. ",
"4A](#fig04){ref-type=\"fig\"} and B; Refs. ",
"\\[[@b48], [@b49]\\]) were used in invadopodia assays.",
"\n\n{#fig04}\n\nFirstly, we confirmed that MMP1--DsRed fusion protein production and matrix degradation can be co-localized. ",
"DQ collagen ([Fig. ",
"4A](#fig04){ref-type=\"fig\"}, green, middle panel) shows matrix underlayment degradation (live cell images) which partially overlaps with MMP1--DsRed production and cellular localization ([Fig. ",
"4A](#fig04){ref-type=\"fig\"}*,* red, middle panel; three representative fields are shown). ",
"The data indicate that MMP1--DsRed expression (red) can be co-localized with degradation (green), creating yellow signals. ",
"Parallel, both can also observe in separate populations in Figure 4A (middle panel).",
"\n\nSecondly, because the DQ-collagen is not MMP-1 specific, we extended this result one step further. ",
"MMP-1 cleavage specific Fluorogenic Substrate-III was additively also incorporated into the underlying gel structures for live cell imaging. ",
"Result in Figure 4B shows that the expressed MMP1--DsRed fluorescent fusion construct (red) co-localized with the strong DQ-collagen degradation signal (green) in the invadopodia (yellow), which was incorporated also the MMP-1 cleavage specific Fluorogenic Substrate-III signal (blue, and white, where all three overlaps). ",
"Effect of pharmacologic inhibition of prenylation on MMP-1 activity (endogenously produced by the cells, no MMP-1 transfection) was examined and verified using only the MMP-1 cleavage specific Fluorogenic Substrate-III ([Fig. ",
"5](#fig05){ref-type=\"fig\"}; 49). ",
"Standardized gel thickness was established as described in the 'Materials and methods' section. ",
"As shown on the live cell images ([Fig. ",
"5A](#fig05){ref-type=\"fig\"}), lightly coloured regions in the untreated control (PlasDIC) correspond to strong MMP-1 enzymatic activity (blue in MMP1-S-III, indicated with white star). ",
"FTI-276 decreased overall MMP-1 activity and eliminated peripheral foci of degradation which is in agreement with the Western blotting results. ",
"Lamellipodia structures in the presence of FTI-276 are associated with less collagenase activity when compared to control or FPP treated cell population (MMP1-S-III, merged [Fig. ",
"5A](#fig05){ref-type=\"fig\"}). ",
"It should be noted that effective invasion depends on lamellipodia-localized MMP-1 enzymatic activity ([Fig. ",
"5B](#fig05){ref-type=\"fig\"}). ",
"Data of invadopodia quantization indicate a 57% activity down-regulation by FTI-276, which was partially restored by FPP with an increase to 75.8% all compared to untreated control podia region (pixel counts 100%).",
"\n\n{#fig05}\n\nDiscussion\n==========\n\nThe current manuscript builds on the previous observations implicating the involvement of MMP-1 in sarcoma metastasis and the prenylation dependence of MMP-1 secretion \\[[@b38],[@b50]--[@b56]\\]. ",
"The results demonstrate that MMP-1 delivery is prenylation dependent and this may affect cellular invasion. ",
"The advantages of performing this study in an established cell line instead of primary sarcoma cells are include but not limited to standardized and uniform gene expressional, invasive and podia formation responses, better support for MetaMorph analysis. ",
"The disadvantage, that established cell line responses seldom reflect the complexity of primary cell populations and could be biased through the immortalization process. ",
"There are two major limitations in this study. ",
"Firstly, the used prenylation inhibition is too broad to determine the detailed molecular mechanism of MMP-1 directional trafficking. ",
"Secondly, the 143B cells over expressing Ras, which in theory might affect the analysis. ",
"Because Ras and FNTA relative quantity did not fluctuate significantly (less than 20% difference; [Fig. ",
"2A](#fig02){ref-type=\"fig\"}) by treatments and we used the Ras only to prove that FTI-276 was working, we are confident that Ras expression did not introduce experimental bias.",
"\n\nIn the absence of a more selective inhibitor of intracellular MMP-1 specific trafficking, prenylation inhibitors represent a convenient way to investigate the subcellular distribution and delivery of MMP-1 to the extracellular space \\[[@b31]\\]. ",
"Interestingly, FPP reversed some, but not all of the changes induced by FTI-276 and did not reverse the cytosolic accumulation of Ras. ",
"This is consistent with only a partial reversal of FNTA activity when examining molecular, morphologic and functional parameters. ",
"The mechanism involving Ras inhibition may or may not be directly related to MMP-1 delivery. ",
"There are reports of Ras regulation of MMP-2 activation and MMP-9 expression which lend support to the linkage between proteases and protein delivery, suggesting an involvement of prenylation \\[[@b57],[@b58]\\]. ",
"Ras-transformed cells are sensitive to FTI-276 as a pharmacologic agent \\[[@b59],[@b60]\\]. ",
"The effect of FTI-276 on MMP-1 gene expression was down-regulated by approximately 30%, which was less than the effect on cellular localization and invasion. ",
"Others have also observed a similar effect in other MMP molecules and in MMP-1, which suggests that additional mechanism may be involved \\[[@b38],[@b61]\\]. ",
"Stable RNAi down-regulation of MMP-1 caused ∼50% decrease in migratory capacity validating its role in directional transmembrane migration ([Fig. ",
"2B](#fig02){ref-type=\"fig\"}--D). ",
"Importantly, the slight up-regulation of MMP-14 (as a result of clonal selection) did not modify MMP-1 knockdown affect on cellular invasion. ",
"The MMP-14 levels gradually decreased in the clones (T6-4 \\> T6-5 \\> T6-7) and this observation did not correlate with the invasiveness patterns ([Fig. ",
"2D](#fig02){ref-type=\"fig\"}).",
"\n\nAn extra limitation of the study is that all determinations were made after overnight incubation while direct prenylation inhibition occurs over the course of minutes to hours. ",
"As such, the relationship between the time course of inhibition and effects on protein trafficking and invasiveness remain unclear. ",
"Previous studies examining the effects of FTI-276 on tumour growth utilized a slow release and a sustained concentration and then documented a 58% reduction in tumour volume over 18 weeks \\[[@b62]\\]. ",
"Abeles *et al*. ",
"measured secretion of MMP-1 from rheumatoid synovium over a 24-hr time period. ",
"Others have reported negative effects of prenylation inhibition at 4 hrs, and statistically significant effects at 24 hrs \\[[@b63]\\]. ",
"Thus, the inhibition kinetics of MMP-1 secretion by FTI-276 is broadly consistent with the known biological processes related to farnesylated proteins. ",
"The fact that farnesylation regulates MMP-1 secretion at least in part at the level of mRNA expression suggests multiple reasons why the effect of FTI-276 on MMP-1 secretion may be delayed.",
"\n\nTo migrate through the collagen-based invasion assay, cells must transverse 8 μm pores and must degrade fibrillar collagen gel in the process (invadopodia related activity). ",
"This report indicates that at least part of the ability to traverse this barrier is MMP-1 dependent and prenylation sensitive. ",
"By inhibiting prenylation, MMP-1 subcellular localization was altered and subsequent collagenase activity and invasion were decreased. ",
"Each of these processes was at least in part reversed by the inclusion of FPP which is an FNTA substrate and served as a prenylation inhibitor agonist \\[[@b61]\\]. ",
"The data demonstrate that MMP-1 directional trafficking and its local activation contributed to the invasion process ([Fig. ",
"4](#fig04){ref-type=\"fig\"}). ",
"It should be noted that (a) not all cells expressed the MMP1--DsRed fusion protein signal (being transiently transformed by the construct); (b) there was more DQ collagen degradation signal than co-localized (could be due to endogenously produced MMP-1 activity or other collagenases); (c) there were also MMP1-DsRed signals that did not show local activation (no green DQ-collagen or blue MMP1-S-III substrate associated with it).",
"\n\nConclusion\n==========\n\nThis paper suggests that MMP-1 directional delivery is an important part of cellular invasiveness and mediated at least in part by prenylation-dependent pathways. ",
"In this regard, directional trafficking is separated from the general secretion process. ",
"The data demonstrates that controlling local lamelli- and invadopodia delivery of MMP-1 through drugs and other complimentary approaches regulate invasive capacity. ",
"Our results illustrate that manipulating directional delivery and can be a beneficial alternative to general metalloproteinase blocking approaches either alone or in combination with other drug treatments.",
"\n\nWe are indebted to Dr. George McNamara and Andras Donaszi-Ivanov at the University of Miami Bio-Imaging Core for their assistance with confocal microscopy and MetaMorph analysis. ",
"We thank Dr. Emily S. Clark for help with the invadopodia assays and David Chyatte, Sara Garamszegi for their contribution to the initial prenylation inhibition data. ",
"This project was supported by ACR 66105H to N.G., NIH CA-66088Y grant to S.P.S. and by the WCA-66461Y grant to N.G. and S.P.S. Other support was provided by the Sylvester Comprehensive Cancer Center and by the University of Miami, Miller School of Medicine.",
"\n\nAuthors' contributions\n======================\n\nN.G. designed the experiments, performed the microscopy data analysis, invadopodia assays, and the final draft of the manuscript. ",
"S.P.G. generated the stable MMP-1 RNAi clones, performed the invasion assays, RT-qPCR amplification, western blotting and related data analysis. ",
"S.P.S. oversaw the collection of data and wrote the first draft of the paper. ",
"All authors read and approved the final manuscript.",
"\n\nConflict of interest\n====================\n\nThe authors have no financial disclosures regarding the performance of the work or the preparation of the current manuscript, and declare no competing and conflict of interests.",
"\n\n[^1]: Present address: 9100 S Dadeland Blvd., ",
"STE 1500 Miami, FL 33156, USA.",
"\n"
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[
"Q:\n\nunderstand the running trace of quicksort in prolog\n\nI have the following prolog code for quicksort:\ngt(X,Y):- X @> Y.\nconc([], List, List).",
"\nconc([Head|Tail], List1, [Head|List2]):- conc(Tail, List1, List2).",
"\n\nquicksort([], []).",
"\nquicksort([X|Tail], Sorted):-\n split(X,Tail,Small,Big),\n quicksort(Small,SortedSmall),\n quicksort(Big, SortedBig),\n conc(SortedSmall, [X|SortedBig], Sorted).",
"\n\nsplit(X,[],[],[]).",
"\nsplit(X,[Y|Tail],[Y|Small],Big):-\n gt(X,Y),\n !,",
"\n split(X,Tail,Small, Big).",
"\n\nsplit(X,[Y|Tail],Small,[Y|Big]):-\n split(X,Tail,Small,Big).",
"\n\nThe array for example is [3,2,4,1,5]. ",
"quicksort([3,2,4,1,5], Sorted). ",
"I need to understand the running trace of the program. ",
"I almost do but there is one point that I don't: (this is just a part of the trace):\nRedo: (11) split(3, [5], _G3785, _G3782) ? ",
"creep\nCall: (12) split(3, [], _G3785, _G3788) ? ",
"creep\nExit: (12) split(3, [], [], []) ? ",
"creep\nExit: (11) split(3, [5], [], [5]) ? ",
"creep\nExit: (10) split(3, [1, 5], [1], [5]) ? ",
"creep\nExit: (9) split(3, [4, 1, 5], [1], [4, 5]) ? ",
"creep\nExit: (8) split(3, [2, 4, 1, 5], [2, 1], [4, 5]) ? ",
"creep\n\nWhy do we have Exit: (11) split(3, [5], [], [5]) ? ",
"creep at the line 11, where does 5 come from?",
"\nSomeone could help me please!! ",
"I'd really appreciate that.",
"\n\nA:\n\nThere should be a cut after comparing at the rule number 2 of function split: \nsplit(X,[Y|Tail],[Y|Small],Big):-\ngt(X,Y), \n!,",
"\nsplit(X,Tail,Small, Big).",
"\n\n"
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[
"Q:\n\nМесто для \"уже\" и \"бы\"\n\nИ второй мой вопрос.",
"\nЯ с тем же примером про \"половину десятого\". ",
"Довольно часто задумываюсь над местом в предложении \"уже\" и \"бы\". ",
"Вроде как, русский язык не запрещает ставить их произвольно, например: \"была уже половина десятого...\" или \"уже была половина десятого...\". ",
"И соответственно: \"Я бы вернулся в половине десятого\", либо \"Я вернулся бы в половине десятого\". ",
"А бывает и так, что в рамках одного предложения частица БЫ встречается вообще несколько раз, например: \"Я бы вернулся в половине десятого, если бы ты заказал мне такси\". ",
"Нормально это? ",
"Или есть возможность избежать повторов частицы бы?",
"\n\nA:\n\nКонечно, такие предложения встречаются. ",
"В классике сотни примеров. ",
"У Льва Толстого их чрезмерно. ",
"И конечно, следует стремиться убирать избыточность, ограничиваясь одним \"бы\". ",
"В вашем примере можно обойтись банальным причастным оборотом: \"Заказав такси, я бы вернулся в половине десятого\". ",
"Либо вообще обойтись без \"бы\": \"Заказав такси, я мог вернуться в половине десятого\".",
"\n\n"
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[
"Exploring Gender Identity\n\nThe Gender Sexual Connection\n\nby Sequoia Elisabeth on June 21, 2011\n\nSex: The physical sexual organs, what is between the legs. ",
"This can change due to sexual reassignment surgery; includes male, female and intersex.",
"\n\nGender: Separate and different from an individual‘s physical sex or sexual orientation (hetero, homo, bi, or asexual), it is an individual‘s perception of the inner self as man, woman or some combination of both, transgender.",
"\n\nOperating from the above definitions we now know that gender and sex are very different and yet from experience we also know that they are intimately connected. ",
"How do we work out this connection? ",
"Is it meaningful?",
"\n\nSexual Orientation is the trait we give the most attention because it involves others. ",
"Who are you attracted to? ",
"Why? ",
"Does gender really have anything to do with this attraction? ",
"How important is appearance? ",
"What about body parts? ",
"Some people are turned on by toes and if this is the case does it matter what is between their legs? ",
"What if ears are a turn on, the same question applies. ",
"You are probably saying that what it leads to is very important and this is where the genitalia come in. ",
"I agree completely.",
"\n\nIn the transgender world we offer a contradiction to this process. ",
"Our parts do not match the majority of the human race. ",
"So do we have GRS to conform or is it more practical than that, simply to enjoy sex in a manner which supports our mental functions? ",
"Many transgender women have surgery just because they cannot bear having male genitalia. ",
"It is like having a large growth on your body which causes you distress. ",
"It could just as well be a tumorous growth! ",
"In this case having surgery is medically necessary, as much as any person with a tumorous growth. ",
"When the result is a happy well adjusted individual post op it is hard to argue against GRS/SRS.",
"\n\n“The reported 68%-86% overall success rates for sex reassignment surgery must be viewed cautiously; the lack of long-term follow-up studies makes these statistics misleading. ",
"There is evidence suggesting that some gender dysphoric patients benefit primarily from sex reassignment surgery. ",
"Most such patients, however, are secondary transsexuals who can benefit from various modes of psychotherapy. ",
"Sex reassignment surgery should only be considered as the last resort for a highly select group of diagnosed gender dysphoric patients. ",
"As physicians learn new ways to diagnose and treat transsexualism, either sex reassignment surgery will be abandoned as a routine treatment modality or new predictive variables for choosing suitable patients for sex reassignment surgery will be established.” (",
"Lothstein, Am J Psychiatry 1982; 139:417-426) The above quote is a bit out dated now and this author finds it interesting that its prediction is far from correct. ",
"SRS has grown in popularity as surgical techniques have been perfected. ",
"It is now the best treatment known for severe gender dysphoria. ",
"The question remains, what if the dysphoria is mild? ",
"What constitutes mild? ",
"This is between the patient and their counselor or therapist.",
"\n\nThe final point or question I wish to raise is, does sexual orientation figure into ones decision to have surgery or not? ",
"From the clinical perspective, the answer would be no. ",
"Dysphoria is separate from sexual behavior and needs to be treated regardless of sexual orientation. ",
"The practical side of the issue is a personal one. ",
"Transsexuals often start off in same sex relationships and to have surgery would destroy the identities associated with the relationship, so unless both individuals are flexible the relationship ends. ",
"This is even the case in heterosexual relationships since it would be homosexual after surgery. ",
"Those relationships that do survive often forgo sex altogether.",
"\n\nOne of the most difficult aspects of the transgender journey is relationships and the changes they must go through along the way. ",
"The losses are often heart breaking, but this is not necessarily a bad thing. ",
"It can lead to more loving and authentic relationships! ",
"The road has many potholes, so it is best to proceed with caution."
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"WTYN\n\nWTYN may refer to:\n\n WTYN (FM), a radio station (91.7 FM) licensed to serve Lunenburg, Massachusetts, United States\n WTYN (AM), a radio station (1160 AM) licensed to serve Tryon, North Carolina, United States, which held the call sign WTYN from 1954 to 1994"
] | {
"pile_set_name": "Wikipedia (en)"
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[
"1. ",
"Field of the Invention\nThe present invention relates to a tuna chunking apparatus and more particularly pertains to allowing a fish to be chunked therein when the tuna chunking apparatus is placed in the operable configuration and further allowing the fish to be chunked by the blades when the projections are forced between the spaces of the blade.",
"\n2. ",
"Description of the Prior Art\nThe use of fish cutting devices is known in the prior art. ",
"More specifically, fish cutting devices heretofore devised and utilized for the purpose of cutting fish into chumb bait are known to consist basically of familiar, expected, and obvious structural configurations, notwithstanding the myriad of designs encompassed by the crowded prior art which has been developed for the fulfillment of countless objectives and requirements.",
"\nBy way of example, U.S. Pat. ",
"No. ",
"5,332,593 to Goto, Tsujii, Okada and Suzuki discloses a method of producing minced fish meat. ",
"U.S. Pat. ",
"No. ",
"5,226,334 to Pegoraro discloses an automatic cutting of meat and fish into portions. ",
"U.S. Pat. ",
"No. ",
"5,205,778 to Korkuch discloses an apparatus for chopping fish for chumb bait and method. ",
"U.S. Pat. ",
"No. ",
"4,794,670 to Savastano, Jr. discloses a fish chunk-cutting device. ",
"U.S. Pat. ",
"No. ",
"4,476,610 to Wenzel discloses a method and apparatus for transversely cutting fish. ",
"Lastly, U.S. Pat. ",
"No. ",
"3,439,721 to Christiansen discloses a machine for cutting bait fish.",
"\nWhile these devices fulfill their respective, particular objectives and requirements, the aforementioned patents do not describe tuna chunking apparatus that allows a tuna chunking apparatus that has a base portion that is positionable on a chum bucket and has a press plate that can be rotated towards the base portion to push the fish through blades of the base portion for chunking of the fish.",
"\nIn this respect, the tuna chunking apparatus according to the present invention substantially departs from the conventional concepts and designs of the prior art, and in doing so provides an apparatus primarily developed for the purpose of the fish to be chunked by the blades when the projections are forced between the spaces of the blade.",
"\nTherefore, it can be appreciated that there exists a continuing need for a new and improved tuna chunking apparatus which can be used for the fish to be chunked by the blades when the projections are forced between the spaces of the blade. ",
"In this regard, the present invention substantially fulfills this need."
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"Review of: Center for Disease Control and Prevention (CDC) Foodborne Outbreak Online Database (FOOD), [wwwn.cdc.gov/foodborneoutbreaks/](wwwn.cdc.gov/foodborneoutbreaks/), CDC Outbreaks, [www.cdc.gov/outbreaks/index.html](www.cdc.gov/outbreaks/index.html), and Multistate Foodborne Outbreaks, [www.cdc.gov/foodsafety/outbreaks/multistate-outbreaks/outbreaks-list.html](www.cdc.gov/foodsafety/outbreaks/multistate-outbreaks/outbreaks-list.html).",
"\n\nEvery season brings a newsworthy outbreak of foodborne infectious diseases such as the 2015 *Escherichia coli* O26 outbreak associated with Chipotle restaurants. ",
"Amid such a nationwide foodborne outbreak, numerous resources can provide students and faculty with timely, accurate, and credible information. ",
"While the Centers for Disease Control and Prevention (CDC) website ([www.cdc.gov](www.cdc.gov)) is a useful starting place, the wealth of information on this website can be overwhelming for new users, such as students. ",
"Two specific resources for understanding foodborne outbreaks include the FOOD (Foodborne Outbreak Online Database) Tool and the Multistate Foodborne Outbreaks webpage. ",
"Both allow users to gain focus when retrieving information on foodborne infectious outbreaks.",
"\n\nThe FOOD tool has been available from the CDC since 2009, but its redesign in 2015 provides a much more user-friendly interface ([@b1-jmbe-17-495b]). ",
"There are multiple entry points into the FOOD tool from the CDC website, the most direct being through [wwwn.cdc.gov/foodborneoutbreaks/](wwwn.cdc.gov/foodborneoutbreaks/). ",
"This is a searchable database of US foodborne outbreaks from 1998 through to 2014, providing at least 16 years of foodborne disease data as of the date of this review ([@b1-jmbe-17-495b]).",
"\n\nOn the entry page of FOOD, the dashboard view provides search option tabs on the left of the page with the remainder of the page displaying a large interactive map, a chart of outbreaks by year, and a chart of outbreaks by month, as well as some brief statistics and other links. ",
"The search option tabs are also very useful. ",
"These tabs are for Year, State, Location of Preparation, Food/Ingredient, and Etiology or causative agent of illness. ",
"Fields can be selected within these search options, and the interactive map, charts and quick stats are automatically updated in real time without delay as search option criteria are selected, added, or removed. ",
"These rapidly generated graphics provide a quick and easy way to understand the quantitative data of any outbreak, which can then be readily communicated to students. ",
"In the same way, students searching for information on a foodborne outbreak can just as rapidly find the information they are looking for.",
"\n\nThe raw data from any searchable outbreak can be very easily downloaded as an Excel data file. ",
"The link for this embedded tool is clearly displayed with the recognizable Excel icon. ",
"This data file contains columns with information on the year, month, state, genus, species, serotype or genotype, etiology status, location of preparation, illnesses, hospitalizations, deaths, food vehicle, and contaminated ingredient. ",
"While some of these data columns mirror the search option tabs, the additional data columns of Serotype or Genotype, Illnesses, Hospitalizations, and Deaths provide a more comprehensive assessment of the outbreak. ",
"In addition, access to this raw data allows users to generate their own tables, graphs, charts and figures. ",
"This is a very helpful teaching tool when generating figures for a lecture but even more so for undergraduate students in introductory microbiology courses who are learning how to access public data, use spreadsheets, generate charts, interpret results, and conduct their own analyses.",
"\n\nOne of the limitations of the FOOD tool is that users are only able to download raw data as Excel files. ",
"There are no direct options to download other file formats such as.csv or .txt, but these file formats can be easily generated from the Excel file. ",
"Unfortunately, an image of the large interactive map cannot be downloaded for repurposing. ",
"There are related minor concerns that are similar, including limited image resolution, the need to create your own state labels, and the inability to change color schemes. ",
"Most importantly, the FOOD database only provides data up to 2014. ",
"With the regular occurrence of foodborne outbreaks, it is expected that this database will be continually updated.",
"\n\nInformation on more recent foodborne outbreaks can be found on the Outbreaks webpage of the CDC's website ([www.cdc.gov/outbreaks/index.html](www.cdc.gov/outbreaks/index.html)). ",
"Conveniently, a link to this webpage is prominently provided on the CDC's homepage. ",
"This link navigates to an informational webpage with a list of current and ongoing \"US-based outbreaks.\" ",
"Each of the listed outbreaks is then hyperlinked to more detailed informational pages. ",
"The \"US-based Outbreaks\" homepage provides a direct link to the \"Multistate Foodborne Outbreaks\" webpage, discussed below. ",
"Other helpful links provided on the \"US-based Outbreaks\" homepage but not discussed in this review include \"Morbidity and Mortality Weekly Reports\" and a widget on \"Food Safety Recalls\" from [www.foodsafety.org](www.foodsafety.org).",
"\n\nThe \"Multistate Foodborne Outbreaks\" webpage ([www.cdc.gov/foodsafety/outbreaks/multistate-outbreaks/outbreaks-list.html](www.cdc.gov/foodsafety/outbreaks/multistate-outbreaks/outbreaks-list.html)) displays information on the most recent 10 years of outbreaks including the two most recent years, which were not included in the FOOD tool. ",
"The information is presented in user-friendly lists and tabs. ",
"There are two sets of lists and tabs: the first set organizes all of the outbreaks by year, while the second set organizes all of the outbreaks by commonly implicated pathogens, such as *Escherichia coli*, *Salmonella*, and *Listeria*. ",
"These lists and tabs make it extremely easy to find information on an outbreak. ",
"For example, by clicking on *2015* and then *Chipotle Mexican Grill*, users are linked directly to a complete informational webpage on the outbreak. ",
"Key statistics on the cases, states involved, hospitalizations, and deaths are quickly gleaned from the 'At a Glance' box. ",
"As well, a clickable map on case counts and a clickable epidemiological curve provide additional details in an easy-to-understand visual format. ",
"Detailed text communicating the 'highlights' of the outbreak and an 'outbreak summary' provide complementary information. ",
"Mid-page clickable tabs with dates provide a timeline of the events that unfolded as the outbreak occurred. ",
"Multiple additional links to general information are also provided. ",
"This Multistate Foodborne Outbreaks resource can facilitate instruction on foodborne outbreaks while complementing content on transmission and pathogenesis. ",
"Faculty, instructors, and students can consult these resources to discuss outbreaks in real time using applied lecture content coupled with accurate information.",
"\n\nEditor's Note: Many instructors use the current newsfeed to provide timely and relatable material to engage students with application-based instruction. ",
"Most printed textbooks lag behind the current news cycle. ",
"In this review, Narveen Jandu, PhD, spotlights the Center for Disease Control and Prevention's database for foodborne outbreaks as one resource instructors and students can access for current, case-based instruction.",
"\n"
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"North Korea has issued a rare apology to a group of South Korean journalists over access to a K-pop concert attended by Kim Jong-un.",
"\n\n\n\nWhile apologies from the North are highly unusual, with criticism often deflected and blamed on the US and its allies, a mea culpa from a senior official over press freedom is almost unimaginable.",
"\n\nThe South Korean reporters had travelled to Pyongyang to cover a K-pop performance, part of a wider rapprochement between the two countries that will see Kim Jong-un meet South Korean president Moon Jae-in later this month.",
"\n\nWhen the journalists were blocked from attending Sunday’s show, the South Korean government lodged an official protest. ",
"Kim Yong-chol, a North Korean general and former spymaster, then went to their hotel to apologise.",
"\n\n“It was wrong to hinder the free media coverage and filming,” Kim Yong-chol said according to South Korea’s Yonhap news agency. “",
"We’re obliged to help reporters from the South side engage in free coverage and convenient filming.”",
"\n\nThe concession on press freedom is very notable and the involvement of Kim Yong-chol, a hardliner blamed by many in the South for an attack that killed 46 navy sailors in 2010, also speaks to the North’s attempts to cool tensions following last year’s nuclear test and missile launches.",
"\n\n“I, representing the North, offer an apology and seek your understanding for the wrong committed,” he added.",
"\n\nHe blamed Kim Jong-un’s bodyguards for blocking media access, saying it was unintentional and caused by miscommunication within the North Korean government. ",
"He also described the event as “special” due to Kim Jong-un’s attendance. ",
"While Sunday’s concert was not the first by South Korean artists in the North, it is the first time the North’s leader has attended.",
"\n\nKim Jong-un “was deeply moved to see our people sincerely acclaiming the performance, deepening the understanding of the popular art of the south side”, according to the North’s official Korean central news agency.",
"\n\nKim met K-pop performers after the concert, shaking each musician’s hand and posing for a group photo. ",
"The scene was at odds with efforts by the North to block South Korean music, often played from loudspeakers on the border as part of the propaganda war between the two neighbours."
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[
"Q:\n\nHow Do I Save This Shirt\n\nThis is the shirt I purchased when I was heavier:\n\nI was able to slim the sides.",
"\nUnfortunately, fixing the shoulders was a train-wreck (now I've learned the hard way: I need to take-in the shoulders)\nWhen I tried to sew along the thread seams, it was crooked and I couldn't put on the shirt. ",
"However, when I tried removing the threads, it made a nice hole in the shoulder area.",
"\nHow to darn this hole so that it is not noticeable?",
"\n\nA:\n\nIt's unlikely that you'll be able to disguise the holes in any way that's not noticeable; instead, I would recommend looking into the Japanese process \"sashiko,\" a mending process made deliberately visible. ",
"Based on your other questions, you are quite new to sewing and embroidery, so you may wish to start with a more simple pattern like the one shown on the bottom; with practice, a wide range of more complex patterns exist that you can use in the future.",
"\n\nA wide range of online tutorials exist to get you started, frequently focusing on a diamond or square pattern similar to the bottom example; the repair will be solid and should help prevent further damage to the area, and will make your shirt uniquely your own.",
"\n\nA:\n\nYou're asking a lot to repair that so it isn't noticeable (although if time or cost is no object it could probably be done; I've seen damaged art canvases invisibly repaired with a microscope, surgical tools, and days of work). ",
" \nA more practical strategy might be to hide it in plain sight. ",
" Incorporate the repair into something like embroidery. ",
" Maybe do something matching on both sides so it looks like it was the original design.",
"\nAnother idea would be to intentionally fray both sides the same way jeans are done.",
"\n\nA:\n\nA less noticable way to repair such a tear is to patch it from behind. ",
"\nCut a patch of matching fabric that covers the whole gap plus one inch in every dimension. ",
"Position the patch behind the tear and hand-sew a few stitches to keep it in place and keep the edges of the tear closed. ",
"You should use a contrasting thread color because you need to remove the stitches later.",
"\nThen choose which stitch and yarn color you want to decorate your shirt with on your sewing machine. ",
"Sew a wild criss-cross pattern all over the tear and make sure the edges of the stitches overlap the edge of the patch below (to keep it from fraying). ",
"I suggest going for a chaotic pattern, because it's very hard to sew a perfectly geometrical criss-cross with a sewing machine and any slight deviation will look odd.",
"\nAt the end, cut and remove the contrasting basting thread.",
"\nThe advantages of this method are:\n\nThe patch isn't visible from the front.",
"\nThe criss-cross stabilizes the fabric in the whole area and reduces the risk of further tearing.",
"\nYou can make it look as if intended.",
"\n\n"
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"INTRODUCTION\n============\n\nCellular senescence is the ultimate and irreversible loss of replicative capacity occurring in primary somatic cell culture. ",
"The discovery of replicative senescence had profound influences not only with respect to the way that ageing is studied, but also how ageing is perceived.",
"\n\nInitially, it was thought that cells once removed from an organism would be able to replicate indefinitely, mainly as a consequence of a long-held claim by Alexei Carrel that chicken embryonic fibroblast cultures could be kept in culture indefinitely ([@B1]). ",
"These findings lead to a widespread notion that ageing was not a consequence of an intrinsic cellular process but some characteristic inherent to the existence of cells in a 'body environment'. ",
"In this context, it is easily understandable how Hayflick\\'s finding of a finite lifespan of human fibroblasts had tremendous impact on our current perception of ageing. ",
"He found that embryo-derived fibroblasts can divide 50 ± 10 times before arresting irreversibly ([@B2]). ",
"This finding suggested that an intrinsic molecular process must account for this phenomenon.",
"\n\nSince then, a thorough characterisation of the various phenotypic changes occurring with senescence had been conducted by various laboratories in the 1970s and 1980s, but none could give unequivocal clues as to the mechanism or mechanisms behind it ([@B3]) until it was suggested that the shortening of telomeres, the ends of chromosomes, could function as a replicometer (counting the finite number of cell divisions) and as a trigger of replicative senescence in normal diploid cells ([@B4],[@B5]). ",
"It was the Russian biologist Alexei Olovnikov who in the late 1960s, after learning about Hayflicks' discovery, first predicted the shortening of telomeres as an explanation for finite cell division in cells grown in culture ([@B6]). ",
"This is still one of the most amazing examples of scientific foresight, since it took more than 20 years to show experimentally that the amount of telomeric DNA does decline with ageing of human fibroblasts ([@B7]). ",
"Of course, it was quite possible that this was a mere marker of senescence like many others that had been observed and no evidence of causality had been demonstrated. ",
"Later, this question was answered by showing that ectopic expression of the catalytic subunit of telomerase, an enzyme able to counteract telomere shortening, can overcome senescence and lead to cell immortalization on its own ([@B8]).",
"\n\nTelomere shortening was proposed as a counting mechanism, which could explain two distinct observations, namely the reproducibility of the 'Hayflick limit' and the fact that cells frozen at a certain population doubling level (PDL) would retain a memory of their PDL and, when thawed, undergo the expected maximum number of divisions ([@B9]). ",
"This was suggestive that a biological programme of ageing was at hand. ",
"The alternative interpretation is that telomere loss is merely a consequence of the cell\\'s inability to synthesise new telomere sequences, and as such, a failure to mobilise resources for maintenance. ",
"As predicted by evolutionary theories of ageing, telomere shortening can thus be seen as an example of limited investment in long-term somatic maintenance and repair function ([@B10]). ",
"There are good reasons why telomere shortening is unlikely to be a counting mechanism. ",
"One is the observation that individual cells from clonally derived populations show heterogeneous division potential ([@B11]) and large heterogeneity in telomere length both between chromosome ends within individual cells and between cells ([@B12; @B13; @B14]). ",
"Moreover, it has been shown that the fraction of senescent cells present in a mass population increases progressively with population doublings, using BrdU labelling ([@B15]), Ki67 staining ([@B16]), p53-reporter assay ([@B17]) and staining for γ-H2AX, a marker for senescence-associated DNA damage foci ([@B18]). ",
"These senescent cells in 'young' cultures showed characteristics of mitochondrial dysfunction including high levels of reactive oxygen species (ROS) together with short telomeres and activation of telomere-induced DNA damage signalling ([@B19; @B20; @B21]). ",
"This indicates that the 'Hayflick limit' can only be applied to mass populations of cells, and that the lifespan of an individual cell lineage is not controlled by a defined genetic programme, but governed by stochastic factors upstream of telomere shortening, probably related to oxidative stress. ",
"Therefore, we will now examine in detail how oxidative stress does affect telomeres specifically and which role mitochondria play in this process. ",
"We will also review the evidence suggesting a role for mitochondria, in particular mitochondrial DNA, in the process of cellular senescence.",
"\n\nTELOMERE ATTRITION IS DEPENDENT ON MITOCHONDRIAL FUNCTION\n=========================================================\n\nThe minimal rate of telomere shortening predicted from the end replication problem is ∼3 bp/end/cell division ([@B22]), however, human cells lacking telomerase lose on average 50--300 bp/end/cell division. ",
"How can this be explained? ",
"There is good evidence, both in *Saccharomyces cerevisiae* and in human cells ([@B23; @B24; @B25; @B26]) that telomere ends are processed by multiple nucleases to generate protruding 3′ ends as part of telomere structure. ",
"To what extent this end processing actually contributes to telomere shortening is less clear. ",
"The simplest idea of a direct correlation between overhang length and telomere shortening rate has been refused experimentally. ",
"A study involving fibroblast strains from 21 donors with 2 orders of magnitude of variation in telomere shortening rate failed to show any correlation between telomere overhang length and shortening rate, suggesting that overhang length does not correlate with telomere shortening ([@B27]).",
"\n\nOxidative stress is another factor that contributes to telomere attrition ([@B28]). ",
"In normal cell culture conditions, cells are exposed to above-physiological levels of oxygen. ",
"While oxygen pressure in the environment is 137 mmHg (corresponding to 21% oxygen at sea level), cells in an organism are exposed to an oxygen partial pressure typically between 3 and 7%. ",
"Moreover, it has been demonstrated that oxygen pressure has a significant impact on the cells' replicative lifespan. ",
"Packer and Fuehr have shown that the replicative lifespan of human diploid cells can be extended by growing them in low oxygen ([@B29]). ",
"Also, by growing telomerase-positive mice embryonic fibroblasts at 3% oxygen for at least 60 days ([@B30]), it has been shown that telomere-independent senescence at the level of a whole culture can be significantly postponed by lowering oxidative stress. ",
"However, these experiments do not exclude the possibility that individual mouse cells grown under physiological oxygen may accumulate sufficient DNA damage to become arrested ([@B31]).",
"\n\nOxidative stress can induce various types of DNA damage, including oxidized bases, single- and double-strand breaks (SSBs and DSBs). ",
"DSBs trigger a DNA damage response that, if persistent, can activate senescence via the p53 and p21 tumour suppressors. ",
"This pathway has been well characterized ([@B32]). ",
"ATM and ATR are recruited to the site of damage and are activated, leading to phosphorylation of the tail of a histone protein variant called 'H2A.X' adjacent to the site of DNA damage. ",
"It is thought that this phosphorylation of histone H2A.X facilitates the focal assembly of checkpoint and DNA repair factors including 53BP1, MDC1/NFBD1 and Nbs1, and also promotes the activation by phosphorylation of Chk1 and Chk2, which converge the signal on p53. ",
"This response occurs when non-telomeric DNA damage is generated by various agents like oxidative stress and ionizing radiation and can lead to induction of a senescent phenotype ([@B33; @B34; @B35]).",
"\n\nTelomeres are normally in a 'capped' state, i.e. unrecognizable to DNA damage response and repair enzyme complexes ([@B36]). ",
"Structurally, they form terminal loops, which are stabilised by a number of telomere binding proteins. ",
"Two of these proteins bind to double-stranded telomeric DNA (TRF1 and -2) and POT-1 binds to single-stranded telomeric DNA. ",
"This complex of proteins has been called 'shelterin', since it 'shelters' i.e. protects chromosome ends ([@B37]). ",
"It is believed that telomere shortening destabilizes telomeric loops ([@B38]) and as a consequence increases the probability of telomere uncapping. ",
"Few years ago, it was shown that uncapping of telomeres, whether by inhibition of TRF2 or telomere shortening, activates the same DNA damage response as do DSBs ([@B18],[@B39]) ([Figure 1](#F1){ref-type=\"fig\"}). ",
"Figure 1.Telomeres shorten with cell division due to the 'end-replication problem' and single-strand break accumulation due to damage by ROS generated as by-product of mitochondrial respiration. ",
"This induces a DNA damage response including formation of telomeric DNA damage foci and activation of p53. ",
"Activated p53 triggers senescent growth arrest.",
"\n\nThus, cellular senescence can be induced as a response to either persistent DSB (telomere-independent or stress-induced premature senescence) or uncapped telomeres (replicative senescence). ",
"Telomere-dependent, replicative senescence is not stress independent, however. ",
"The intensity and dosage of oxidative stress determine the probability of DSB generation. ",
"Intense, acute stress will generate DSBs at higher frequency and might modify these broken DNA ends to make them more resistant to repair, leading to DSB persistence and so induction of senescence via non-telomeric DNA damage response. ",
"Chronic oxidative stress of low intensity mainly generates oxidative base modifications and base excision repair intermediates, i.e. abasic sites and SSBs.",
"\n\nTelomeres acquire such oxidative single-strand damage faster than the bulk of the genome for two reasons: First, sequences containing guanine triplets are exquisitely sensitive to oxidative modification ([@B40],[@B41]). ",
"For instance, a human telomere sequence inserted in a plasmid showed up to 7-fold more strand breakage than a control sequence ([@B41]). ",
"Second, repair of SSB ([@B42]), and to some extent also of UV-induced damage ([@B43]), is significantly less efficient in telomeres as compared to the bulk of the genome. ",
"This holds true even for a comparison between telomeres and interstitial guanine-rich repetitive sequence tracts ([@B42]). ",
"The cause for this telomere-specific repair deficiency has not been established with certainty yet. ",
"However, a structural basis for it is probable, since TRF2 binds to telomeric double-stranded DNA, stabilizes the telomeric loop and thus contributes to telomere capping. ",
"Overexpression of TRF2 decreases the telomere length threshold at which replicative senescence is signalled in accordance with improved capping ([@B44]). ",
"However, it also accelerates the rate of telomere shortening ([@B44]) and further decreases the efficiency of SSB repair specifically in telomeres ([@B45]). ",
"The most simplistic interpretation of these data is that telomere capping would be detrimental to free access of DNA repair complexes to telomeres. ",
"It has been shown that TRF2 (and TRF1) binding can stall the replication fork progression at telomeres ([@B46]). ",
"However, it needs to be mentioned that TRF2 plays more complex roles at telomeres as it interacts with and inhibits enzymes involved in DNA repair including ATM ([@B47]) and Polymerase β ([@B48]), while its binding to telomeres is reduced by oxidative DNA damage ([@B49]).",
"\n\nIrrespective of the specific mechanism causing repair inefficiency, telomeres enter DNA replication with higher frequencies of single-strand DNA damage than all the bulk of the genome. ",
"This contributes significantly to telomere shortening ([@B50]), with the frequency of single-strand damage directly correlated with the amount of telomere loss ([@B51]). ",
"Oxidative damage-dependent telomere shortening might be caused by a temporal stalling of the replication fork or might be recombination dependent ([@B52]). ",
"It might be a significant cause of telomere length heterogeneity between different chromosomes and between the same chromosomes in different clonally derived cells ([@B12],[@B19]).",
"\n\nFinally, oxidative stress interferes with telomere maintenance also via its effect on telomerase activity. ",
"Telomerase is a ribonucleoprotein complex that consists of two main components -- the RNA component (TER), containing the antisense template sequence for telomere synthesis and the catalytic protein, telomerase reverse transcriptase --(TERT). ",
"Telomerase re-elongates telomeres in a highly regulated fashion to counteract telomere shortening. ",
"This is sufficient to confer extended lifespan and often immortality to cells that would otherwise senesce due to continuous telomere shortening ([@B8]). ",
"Physiologically, human telomerase is expressed in germ line and stem cells and in most cancers, but its expression is low (haematopoetic cells and endothelial cells) or undetectable without sophisticated enrichment protocols ([@B53]) in somatic human cells like fibroblasts. ",
"Oxidative stress diminished telomerase activity in endothelial cells ([@B54; @B55; @B56]), vascular smooth muscle cells ([@B57]) or leukemic cancer cell lines ([@B58]) together with an acceleration of telomere attrition. ",
"However, oxidative stress had no effect on telomerase activity in fibroblasts overexpressing TERT ectopically ([@B19]), suggesting preferentially transcriptional regulation.",
"\n\nTwo properties of oxidative stress-dependent telomere attrition are important. ",
"First, it is dependent on DNA replication. ",
"The probability of generating DSBs directly at a frequency high enough to result in a measurable telomere loss without DNA replication is negligible for physiologically and pathologically obtainable stress levels. ",
"Second, oxidative damage can be a major, and often actually the main, contributor to telomere loss in cell culture under standard conditions. ",
"This is shown by the fact that telomere shortening rates in cultured human fibroblasts not only accelerate if oxidative stress is increased, but that they can be diminished if oxidative stress levels are reduced below those in standard conditions by using free radical scavengers ([@B59]), enzymatic ([@B60]) or non-enzymatic ([@B61]) antioxidants or low ambient oxygen concentrations ([@B62],[@B63]), in some cases to levels similar to the minimal values predicted by the end replication problem alone. ",
"For instance, we measured telomere shortening rates of 7 ± 9 bp/PD in BJ fibroblasts, a strain with very high antioxidant capacity ([@B28],[@B60]), and of 9 ± 29 bp/PD in MRC5 fibroblasts treated with Dinitrophenol, an agent that reduces mitochondrial ROS generation by mild uncoupling ([@B20])\n\nNow, the important question is: Is oxidative stress-dependent telomere length regulation merely a response to more or less artificial changes in cell culture environments (i.e. a 'cell culture artefact')? ",
"or is it part of an intrinsic mechanism governing the ageing process? ",
"Correlative evidence from human population studies collectively suggests an association of short telomeres with conditions of increased oxidative stress, including smoking, obesity, various cardiovascular diseases, psychological and socio-economic stress ([@B64; @B65; @B66]). ",
"Cells in telomere-dependent senescence accumulate in aging baboon skin ([@B67],[@B68]). ",
"We propose mitochondrial dysfunction as a major mechanistic link between stress-dependent and telomere-dependent physiological ageing processes in cells. ",
"Mitochondria are the major source of oxygen-free radicals in cells. ",
"It has been revealed that mitochondrial function changes as cells reach the end of their replicative lifespan, leading to increased generation of ROS and metabolic inefficiency ([@B20],[@B69; @B70; @B71; @B72]). ",
"Increased ROS leads to accumulation of oxidation products, such as protein carbonyls and lipofuscin, which have been shown to occur in senescent fibroblasts ([@B73],[@B74]).",
"\n\nIn addition to correlative evidence showing mitochondrial dysfunction in telomere-dependent senescence, there is also good evidence supporting a causal role for mitochondrial dysfunction in the process. ",
"Selective targeting of antioxidants directly to the mitochondria has been shown to counteract telomere shortening and increase lifespan in fibroblasts under mild oxidative stress ([@B61]). ",
"Continuous treatment with nicotinamide, which affects mitochondrial function and ROS generation, has been reported to extend lifespan (an amazing 1.6-fold increase) and decelerate telomere shortening ([@B75]). ",
"Also, mild chronic uncoupling of mitochondria that reduced the production of superoxide anion, improved telomere maintenance and extended telomere-dependent lifespan ([@B20]).",
"\n\nOn the other hand, mitochondrial dysfunction generated by severe mitochondrial depolarization using an uncoupling agent FCCP led to an increased production of ROS, telomere loss and chromosome fusions in mouse embryos ([@B76]). ",
"Moreover, Oexle and Zwirner showed that patients with mitochondrial diseases MELAS and LHON had, on average, 1.5 kb shorter telomeres in white blood cells than those of age-matched controls ([@B77]).",
"\n\nThis set of data is suggestive of a novel role for telomeres as sensors of mitochondrial function in a cell ([@B78]). ",
"When mitochondrial dysfunction occurs, with concomitant increased ROS generation, the specific susceptibility of telomeres to oxidative damage ([@B42]) leads to accelerated telomere shortening, increased probability of uncapping, activation of a DNA damage response and finally irreversible cell cycle arrest ([Figure 1](#F1){ref-type=\"fig\"}). ",
"Thus, as previously suggested, the replicative lifespan of a cell could be determined by a network of processes involving mitochondrial dysfunction, oxidative stress and telomere shortening ([@B20],[@B78],[@B79]).",
"\n\nHowever, one important question remains: What is the cause of mitochondrial dysfunction and ROS generation? ",
"Is it dependent on damage to the mitochondrial genome?",
"\n\nMTDNA DAMAGE AND SENESCENCE: IS THERE A LINK?",
"\n=============================================\n\nMitochondrial DNA has for a long time been implicated in the ageing process. ",
"The idea was first proposed by Harman, who predicted the involvement of free radicals in the ageing process ([@B80],[@B81]). ",
"The concept was that the close proximity between the sites of ROS production and the mtDNA would render the latter more susceptible to damage than the nuclear genome, and lead to defects in mitochondrial metabolism. ",
"The long-held view that the mitochondrial genome is not protected by histones and mtDNA repair is inefficient has been challenged in recent years by showing that TFAM acts like a histone covering mtDNA ([@B82]) and that there exist functional repair mechanisms for different types of DNA damage within mitochondria with the possible exception of nucleotide excision repair ([@B83]). ",
"Even so, it has been shown that mtDNA damage is more extensive and persistent than nuclear DNA damage in cultured human fibroblasts following treatment with hydrogen peroxide ([@B84]) and this correlates with functional impairment in mitochondria ([@B85]). ",
"Treatment of normal human fibroblasts with sublethal levels of oxidative stress has also been associated with accumulation of the mtDNA common deletion, a specific 4977 bp deletion often found in mtDNA ([@B33]).",
"\n\nAn age-dependent increase of mtDNA mutation frequency was first observed in post-mitotic cells such as neurons and muscle cells ([@B86; @B87; @B88; @B89]), but has since been described in highly proliferative cells such as the epithelial stem cells of the gut wall as well ([@B90]). ",
"mtDNA mutations are responsible for deficient activity of respiratory chain enzymes such as Cytochrome C oxidase, which is a mitochondrial membrane--bound enzyme composed of subunits that are encoded in both the mitochondria (COX subunits I, II and III) and the nucleus (all others). ",
"The frequency of COX-deficient human heart muscle cells was shown to increase significantly with age, but remained always below 1% ([@B88]). *",
"In situ* hybridization studies using mtDNA probes showed accumulation of mtDNA deletions in COX negative cells ([@B91]). ",
"Using microdissection to isolate muscle fibres, it was shown that all electron transport chain--deficient fibres in rat skeletal muscle contained mtDNA deletion mutations ([@B92],[@B93]). ",
"In single neurons from *substantia nigra* of humans showing COX deficiency, an age-dependent increase in mtDNA deletions was found in two independent studies ([@B94],[@B95]).",
"\n\nConsequently, an attractive hypothesis is that the increase of mutational load in mtDNA with age contributes causally to mitochondrial dysfunction and increased ROS production, so creating a *circulus vitiosus*. ",
"However, new data question the causal link between mtDNA mutations and ROS generation. ",
"Homozygous knock-in mice expressing a proof reading--deficient version of the nucleus-encoded catalytic subunit of mtDNA polymerase γ (PolgA) have been generated that showed an extremely high level of mtDNA mutations and deletions and a significant decrease in lifespan ([@B96]). ",
"However, even though mitochondrial function was affected, no evidence for increased oxidative stress was found in these animals ([@B97],[@B98]). ",
"Most strikingly, mice that are heterozygous for PolgA function showed no significant reduction in lifespan despite a mtDNA point mutation burden 30 times higher than in old wild-type animals ([@B99]). ",
"These studies suggested that mtDNA point mutation load does not limit lifespan of wild-type mice and that mtDNA point mutations, even at very high levels, do not necessarily lead to increased mitochondrial ROS generation. ",
"It remains to be seen whether there is a more sinister role for mtDNA deletions in aging.",
"\n\nSo far, there is very little data to assess the role, if any, of mtDNA mutations in cellular replicative senescence. ",
"Accumulation of mtDNA point mutations in fibroblasts isolated from old individuals has been reported ([@B100]). ",
"Most strikingly, a T414G transversion was found in a high proportion (up to 50%) of mtDNA molecules from individuals above 65 years of age. ",
"However, when several fibroblasts populations carrying the heteroplasmic T414G mutation were grown *in vitro*, outgrowth of the mutant cells by wild-type cells was observed ([@B101]). ",
"This led to the suggestion that accumulation of mtDNA point mutations is a phenomenon occurring exclusively *in vivo.*",
"\n\nUsing a real-time PCR-based method ([@B102]), we have recently found evidence of increased mtDNA damage in senescent human fibroblasts ([@B20]). ",
"However, these are correlative data and it is not possible to discern whether mtDNA damage caused mitochondrial dysfunction or was a consequence of it. ",
"It could well be that other factors besides mtDNA damage and/or mutation might be more important for the generation of ROS. ",
"Such factors could include the regulation of mitochondrial turnover or mitochondrial fusion and fission ([Figure 2](#F2){ref-type=\"fig\"}) in addition to extra-mitochondrial ones. ",
"Figure 2.Possible mechanisms influencing mitochondrial ROS generation: (1) mtDNA damage could lead to dysfunctional respiratory chain activity and increased ROS, (2) mitochondrial elongation by decreased fission has been shown to increase ROS generation, (3) increased mitochondrial biogenesis has been shown to correlate with ROS generation and induce cellular senescence. ",
"ROS-generated telomere shortening as well as double-stranded breaks at non-telomeric DNA activate a DNA damage response and cellular senescence.",
"\n\nAccordingly, a recent study has shown that mitochondrial elongation by knock down of the mitochondrial fission protein hFis1 led to induction of senescence, possibly through increased ROS generation and activation of a DNA damage response ([@B103]). ",
"Moreover, there is some evidence suggesting that mitochondrial ROS generation in senescence might be due to increased mitochondrial density, which can occur as a consequence of mitochondrial dysfunction ([@B20],[@B104]). ",
"In fact, it has been shown that overexpression of PGC-1α, the master regulator of mitochondrial biogenesis, leads to induction of senescence in human fibroblasts ([@B105]). ",
"The association between increased mitochondrial density and ROS generation might be due to amplification of damaged mtDNA molecules. ",
"In summary, the role of mtDNA damage and mutation in senescence is still unclear.",
"\n\nTelomerase and oxidative stress: a role beyond telomeres\n--------------------------------------------------------\n\nIn recent years, indications for additional functions of telomerase independent of telomere maintenance are accumulating. ",
"If telomerase is inhibited in tumour cells, there is either a delayed response that depends on telomere shortening or a rapid effect on cell viability without any measurable effect on telomere length ([@B106; @B107; @B108; @B109; @B110]). ",
"These data suggest that telomerase promotes cell survival and stress resistance by mechanisms that appear to be largely independent on telomere length maintenance. ",
"Zhang *et al.* ",
"and Fu *et al.* ([",
"@B111],[@B112]) showed an increased resistance of hTERT overexpressing cells to apoptotic stimuli at an early, premitochondrial step. ",
"Sharma *et al.* ([",
"@B113]) found an increased repair capacity of hTERT overexpressing cells. ",
"We and others demonstrated an enhanced sensitivity of tumour cells to certain DNA damaging agents when telomerase was impaired ([@B107],[@B113; @B114; @B115; @B116; @B117]). ",
"We showed that overexpression of TERT conferred increased stress resistance, improved antioxidant defence and differentiation capacity to mouse embryonic stem cells ([@B118]).",
"\n\nEctopic hTERT expression in normal cells as well as hTERT inhibition/depletion in telomerase positive cancer cells and even in yeast *S. cerevisiae* caused significant changes to the transcriptome and global gene expression patterns ([@B106],[@B108],[@B119],[@B120]) that are largely unrelated to telomere maintenance. ",
"Interestingly, among a wide range of functional classes, large groups of genes with functions in metabolism, specifically mitochondrial metabolism, have repeatedly been reported to appear dependent of telomerase ([@B106],[@B119]). ",
"Bagheri *et al.* ([",
"@B106]) found that telomerase modulated glucose consumption and appeared to control the glycolytic pathway in tumour cells, thereby potentially altering the energy state. ",
"Recently, telomerase has been added to a growing list of proteins, such as p53, HMGA1, VHL, APP, prohibitins, Lon-protease, etc., ",
"that can shuttle between the nucleus and the different subcellular compartments including mitochondria. ",
"Importantly, it has been shown that telomerase that normally is located in the nucleus can shuttle to cytoplasm and/or mitochondria upon oxidative challenge ([@B55],[@B121; @B122; @B123]).",
"\n\nSantos *et al.* ([",
"@B117]) described a specific mitochondrial import sequence at the N-terminus of hTERT. ",
"Oxidative stress activates nuclear export and mitochondrial import for both ectopically expressed and endogenous telomerase ([@B55],[@B121; @B122; @B123]). ",
"Haendeler *et al.* ([",
"@B55]) found that nuclear export of telomerase occurs during *in vitro* senescence of human endothelial cells and can be delayed by treatment with antioxidants. ",
"One possible mechanism connecting oxidative stress and nuclear exclusion of telomerase is the activation of Src kinases by ROS ([@B124]). ",
"It has been shown that phosphorylation of tyrosine 707 by Src kinase is necessary for nuclear exclusion of hTERT following oxidative stress ([@B121]). ",
"A similar process has been described for T-lymphocytes where telomerase function is regulated via phosphorylation and nuclear translocation ([@B125]). ",
"These data suggest that subcellular shuttling is not an artefact of ectopic hTERT expression. ",
"Rather, transport of hTERT to mitochondria is a directed, naturally occurring process that is regulated by either exogeneously or endogenously generated oxidative stress.",
"\n\nThe biological significance of the translocation of hTERT into mitochondria under oxidative stress is still elusive. ",
"Santos *et al.* ([",
"@B122],[@B123]) found that ectopic overexpression of hTERT led to higher levels of mtDNA damage under acute oxidative stress. ",
"In contrast, there is an increasing number of papers showing the telomerase protects mitochondrial function and displays an anti-apoptotic function. ",
"Massard *et al.* ([",
"@B109]) characterised hTERT as an endogenous inhibitor of mitochondrial apoptosis induced by different agents including oxidative stress in cancer cells. ",
"Neurons with decreased hTERT levels exhibited increased levels of oxidative stress and mitochondrial dysfunction after exposure to amyloid beta, whereas overexpression of hTERT in the same system led to an improved mitochondrial function and a decreased oxidative stress level ([@B126]). ",
"Kang *et al.* ([",
"@B127]) found that after ischemic brain injury TERT is induced in postmitotic neurons in TERT transgenic mice and prevents NMDA neurotoxicity via shifts of free cytosolic free Ca^2+^ to the mitochondria. ",
"The authors found an enhanced basal level of mitochondrial membrane potential and a higher Ca^2+^ storage capacity of the mitochondria due to TERT overexpression. ",
"Our own data indicate protection of mtDNA and mitochondrial function under oxidative stress by ectopically expressed hTERT (unpublished).",
"\n\nThese results show that there is an interaction between mitochondrially localised hTERT and mitochondrial function that seems to be complex and can involve different physiological and signalling pathways. ",
"It is not yet clear whether mitochondrial localisation of hTERT is necessary to improve mitochondrial function and/or to protect cells from stress-induced apoptosis.",
"\n\nCONCLUSIONS\n===========\n\nIt appears highly probable that damage to two specific subsets of cellular DNA, namely telomeres and mtDNA, plays an important role in cellular senescence. ",
"These two types of DNA damage are functionally interrelated at various levels. ",
"mtDNA damage, especially deletions, might contribute to mitochondrial dysfunction and ensuing ROS production, which is a major causal factor for telomere damage and shortening, resulting eventually in senescence signalling. ",
"Conversely telomerase, the central enzyme in telomere length maintenance, can translocate to mitochondria under stress and impacts on mtDNA protection and mitochondrial function. ",
"It is possible that there is a self-amplifying cycle between mitochondrial and telomeric DNA damage driving cellular senescence.",
"\n\nWork leading to this review was supported by programme grant 252 from Research into Ageing, UK and by a BBSRC/EPSRC systems biology grant (CISBAN). ",
"Funding to pay the Open Access publication charges for this article was provided by BBSRC.",
"\n\n*Conflict of interest statement*. ",
"None declared.",
"\n"
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[
"Brachytherapy is a type of radiation therapy used to treat malignant tumors, such as cancer of the breast or prostate. ",
"In general, brachytherapy involves positioning a radiation source directly into target tissue, which may include a tumor and/or tissue surrounding a cavity or void, which may contain potentially cancerous cells (such as a cavity or void created by removing a tumor).",
"\nBrachytherapy is often divided into two categories: high dose rate (HDR) and low dose rate (LDR) brachytherapy. ",
"In HDR brachytherapy, a high activity radiation source is placed into target tissue, often via a previously implanted catheter, for a short period of time, e.g., lasting from several seconds to a few minutes. ",
"In contrast, LDR brachytherapy involves placing a low activity radiation source into the target tissue for a longer, sometimes indefinite, period of time.",
"\nBoth forms of brachytherapy have advantages. ",
"For instance, HDR brachytherapy provides higher radiation levels delivered over a shorter dose delivery period. ",
"LDR brachytherapy, on the other hand, utilizes lower activity radiation sources. ",
"The energy field of the LDR radiation source results in a measured and localized dose of radiation delivered to target tissue, e.g., a tumor, gland, or other tissue surrounding a cavity or void. ",
"However, the energy field thereafter decays to avoid excessive exposure of nearby healthy tissue.",
"\nDue in part to the lower activity of LDR radiation sources, LDR brachytherapy may provide various advantages. ",
"For example, for healthcare workers, exposure precautions for LDR brachytherapy may be less stringent than those for HDR brachytherapy. ",
"Also, there are radiobiological advantages of LDR brachytherapy over HDR brachytherapy (e.g., the dose rate effect), which may lead to better sparing of normal tissue during treatment. ",
"Moreover, for patients, the relatively longer implantation period associated with LDR brachytherapy may result in fewer visits to a healthcare facility over the course of radiation treatment, as compared to HDR brachytherapy where patients must return to the healthcare facility for each fraction of radiation delivered, which, for breast brachytherapy, may typically include eight to ten (8-10) fractions.",
"\nWhile effective, current brachytherapy implementations have potential drawbacks. ",
"For example, LDR seeds are typically left indwelling and free floating within the target tissue and are, therefore, susceptible to migration. ",
"Moreover, once implanted, LDR seeds are generally not considered removable or repositionable. ",
"LDR brachytherapy may also require careful dose distribution calculations and seed mapping before, and often during, seed implantation. ",
"Such calculation and mapping may allow effective radiation delivery to the target tissue volume, while minimizing radiation to surrounding healthy tissue (e.g., the urethra and rectum, for example, in prostate brachytherapy). ",
"Yet, while such dose calculation and seed mapping techniques are effective, problems may exist, such as potentially significant variability in accuracy of seed placement among different clinicians.",
"\nYet another issue with conventional LDR brachytherapy techniques is that they may require the radioactive seeds to be manipulated individually at the time of implantation, which may be a time-consuming process. ",
"Moreover, conventional LDR delivery needles are generally limited to delivering the seeds linearly (along a relatively straight line). ",
"Thus, to achieve a desired therapy profile, numerous implants (e.g., including about 50-100 seeds, as are common with prostate brachytherapy) are often required, in conjunction with potentially complex dose distribution and mapping techniques and equipment."
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[
"I'm trying APV Beta version, when i render the file i get a file of only 5 seconds. ",
"Is this part of the trial version limitations? ",
"or i'm doing something wrong? ",
"i've already worked with beta 3 version and i wasn't having this problem.regards.gon\n\nThank you very much Georg. ",
"I wasn't looking that deep at the documentation.",
"\n\nSo, here we go again:Cannot get a rendered video longer than 5secs. ",
"All test videos are around 57sec, default settings from go pro.",
"I've already test it once with beta version 3 and were ok, now i improved the stitch and i want to try it again."
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"William Hunden\n\nWilliam Hunden was Archdeacon of Totnes from 1408 until 1415.",
"\n\nReferences\n\nCategory:Archdeacons of Totnes\nCategory:Medieval English clergy"
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[
"Q:\n\nConditional Calucations with MVC4\n\nI am trying to conditionally calculate some values based on user inputs in a form. ",
"The \"inputs\" themselves tie into the calculation, in that if a user is presented with two radio button lists their selections will determine whether or not a currency amount is attached, and then later added together. ",
"I don't understand jquery, and so I cannot figure out how to use something like knockout.js to do what I want (besides, my calculation will be served up in a confirmation view only and will not be \"re-calculated\" unless the user goes back and changes their selections).",
"\nNote, I am not talking about the scenario where you have a textbox that accepts a value and adding it to another textbox value the user inputs (e.g., user enters 10, then 10, then form calculates 20). ",
"The values are static based upon selection.",
"\nFor context, I am using Serializer and a wizard which is stepping through and passing model selections/inputs to the subsequent view.",
"\nI can do something like this as an example (it's simple, but based on responses I can figure out how to build it up to the more complex scenario I have):\nModel.cs:\n[DataType(DataType.",
"Currency)]\n[DisplayFormat(DataFormatString = \"{0:c}\")]\npublic decimal calculated { get; set; }\n\n[DataType(DataType.",
"Currency)]\n[DisplayFormat(DataFormatString = \"{0:c}\")]\npublic decimal option1 = 500;\n\n[DataType(DataType.",
"Currency)]\n[DisplayFormat(DataFormatString = \"{0:c}\")]\npublic decimal option2 = 100;\n\n//Following is just my idea to calculate $0 if \n//a selection results in a \"no value option\"\n//although I suppose if nothing is selected I \n//can just avoid calculating + $0\n[DataType(DataType.",
"Currency)]\n[DisplayFormat(DataFormatString = \"{0:c}\")]\npublic decimal nooption = 0;\n\nThen in Confirm.cshtml:\n@{\n Model.calculated = Model.option1 + Model.option2 + Model.nooption;\n}\n....\n@Html.",
"DisplayFor(m => m.calculated)\n\nwill display $600.00. ",
"That's easy enough.",
"\nHowever, my goal is threefold. ",
"First, I need to tie in, for example, option1 with the selection from a radiobutton list, and option2 with another radiobutton list depending on what the user selects. ",
"Second, depending on the selection I want to conditionally calculate all the options present which will obviously vary based on user selection (some users will select option1 and option2, others just option1, others just option2, still others neither). ",
"Third, where is the best place to have the calculation take place (the view, a .cs file, the controller (although I am trying to minimize code there), etc.).",
"\nSo, if I have these enums for the radiobutton lists (each option corresponding to an option for now, later I will discuss two options per radiobutton list enum):\npublic enum radio1\n{\n radio1selection1, // this would correspond to nooption (or $0)\n radio1selection2 // this would correspond to option1 (or $500)\n}\npublic enum radio2\n{\n radio2selection1, // this would correspond to nooption (or $0)\n radio2selection2 // this would correspond to option2 (or $100)\n}\n\nI've indicated in the enums how I'd like to \"tie\" the option to the selection.",
"\nMy problem is in trying to do if statements, which I cannot figure out properly. ",
"Something like this in Confirm.cshtml:\n@{\n if (Model.radio1 == radio1selection1)\n Model.calculated = Model.nooption;\n else....\n}\n\nThe above is completely wrong, I know. ",
"I am trying to think of all the permutations of calculating, but I don't think its worth the effort because it would either be wrong or it would work but be too long when there is an easier way. ",
"I was even thinking of trying case/break but that too seems oddly the wrong approach.",
"\nWhat's adding to the complexity for me is if my enums have more than two selections, and those other selections result in several options, say option1 and option1a, and depending on which the user selects will determine the calculation. ",
"So the enum becomes:\npublic enum radio1\n{\n radio1selection1, // this would correspond to nooption (or $0)\n radio1selection2, // this would correspond to option1 (or $500)\n radio1selection3 // this would correspond to NEW \n // option1a (or $750) (with a corresponding \n // \"int\" named \"option1a\")\n}\n\nOne of the things I thought of doing was to just display all the options selected in a <table> in the view, so that even if something is a $0 value, it would just say $0 as the option. ",
"Then I thought I could just add all the values present. ",
"But still I don't know the best way to accomplish that.",
"\nAny opinions on how I would be able to and or all three of my stated goals above?",
"\n\nA:\n\nI figured out how to do this (might not be clean/efficient/standard practice, but it got the job done):\n Model.calculated =\n Model.",
"PriceQuote.priceChapter7Solution +\n ((Model.radiobuttonlist1 == \n Namespace.",
"ViewModels.",
"MyModel.radio1.radio1selection1) ?",
"\n Model.option1 :\n (Model.radiobuttonlist1 == \n Namespace.",
"ViewModels.",
"MyModel.radio1.radio1selection2) ?",
"\n Model.option2 :\n Model.nooption);\n\nI could just add + after ... : Model.nooption) instead of ending the statement with ; to calculate conditions on other options.",
"\nOriginally I did not know that the conditional (? :) ",
"operator could contain more than two expressions - i.e., it's right associative so additional conditions will be evaluated as a ? ",
"b : (c ? ",
"d : e). ",
"See Docs.",
"\n\n"
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"This sample Android app demonstrates\nanimating markers in a map.",
"\n\nThis app is covered in \n[the chapter on Maps V2](https://commonsware.com/Android/previews/mapping-with-maps-v2)\nin [*The Busy Coder's Guide to Android Development*](https://commonsware.com/Android/).",
"\n\n"
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