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The 5C's Checklist for Recruiting
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Before You Hire, Check Off This Recruiting Checklist
We all know hiring employees for your business is not an easy task. There are several factors and considerations to think about when finding the perfect candidates. To make your recruiting process a bit smoother, here's a checklist of five characteristics to keep in mind:
1. Competence
This should be the first factor to consider. Does the candidate have the necessary skills, experiences and educational background to succeed? If he/she does not fulfill those requirements, see if he/she is willing to be trained for the position.
2. Compatible
Look to see if this person can get along with colleagues, and most importantly, existing and potential clients. It is a critical component that he/she be harmonious with your company's people and those that may walk through your doors in the future.
3. Commitment
Another question to ask yourself when hiring is, "Is the candidate serious about working for the long term?" If he/she seems like they are not in it for the long run, you most likely want to pass on them. You do not want to go through the hiring process once again in a few months time. You could be using your valuable time, money and effort on more important things.
4. Character
Does the person have values that align with yours? Is he/she honest, keep promises, or is a team player? Do consider these things when making your final decision because you certainly do not want a bad apple to spoil the whole bunch (the company).
5. Culture
Every business has a culture or a way that people behave and interact with each other. The company culture is based on values, expectations, policies and procedures that influence the behavior of a leader and employees. If he/she doesn't reflect the company's culture, he/she may be disruptive and difficult to work with.
About the Author
Christie, currently a editorial assistant at Harri, enjoys exploring and eating her way through her hometown of New York City.
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- There are several factors and considerations to think about when finding the perfect candidates.
- Does the candidate have the necessary skills, experiences and educational background to succeed?
- Does the candidate seem like they would fit in and get along with your the current employees?
- Is this candidate a team player and looking to stay with your company for the long-term?


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Die Entwicklung des Klavierkonzerts von Liszt bis zur Gegenwart1 edition - first published in 1931
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TABASCO: A single molecule, base-pair resolved gene expression simulator
Experimental studies of gene expression have identified some of the individual molecular components and elementary reactions that comprise and control cellular behavior. Given our current understanding of gene expression, and the goals of biotechnology research, both scientists and engineers would benefit from detailed simulators that can explicitly compute genome-wide expression levels as a function of individual molecular events, including the activities and interactions of molecules on DNA at single base pair resolution. However, for practical reasons including computational tractability, available simulators have not been able to represent genome-scale models of gene expression at this level of detail.
Here we develop a simulator, TABASCO
Tabasco enables genome-scale simulation of transcription and translation at individual molecule and single base-pair resolution. By directly representing the position and activity of individual molecules on DNA, Tabasco can directly test the effects of detailed molecular processes on system-wide gene expression. Tabasco would also be useful for studying the complex regulatory mechanisms controlling eukaryotic gene expression. The computational engine underlying Tabasco could also be adapted to represent other types of processive systems in which individual reaction events are organized across a single spatial dimension (e.g., polysaccharide synthesis).
Mechanistic models of the individual biochemical events that comprise gene expression are quite detailed and are continuously improving. For example, experiments studying the kinetics of promoter initiation and single molecule studies of processive protein movements along DNA have revealed intricate regulatory processes that control gene expression
There are several mathematical approaches for computing the dynamics of systems of biochemical reactions. For example, the chemical species that define a system can be modeled as continuous variables that change over time. In this "continuous" approach, reactions between chemical species are modeled as a set of coupled ordinary differential equations (ODEs)
However, challenges exist in using either approach to study high-resolution models of gene expression. Both discrete and continuous approaches to solving the time evolution of biochemical reactions share the so-called "combinatorial explosion" problem, in which the number of possible states that need to be enumerated in a system becomes exceedingly large
Two approaches have addressed the combinatorial explosion problem with respect to protein complex formation and modification during signaling
StochSim and Moleculizer were designed to model the binding and unbinding of proteins within complexes. However, these simulators are not well suited for modeling systems in which components within a complex are processive (i.e., components transitioning among many binding states) or when the number of binding states is large. For example, simulating a single RNA polymerase transcribing a 3,000 bp gene at single-base resolution using a single-molecule simulator would require a model for a DNA molecule comprised of over 3,000 different states with enumerated rules for the transitions from one base to the next. Using a dynamic reaction generation simulator, such as Moleculizer, would not lead to significant speed enhancements because most of the states of the system are reached during a simulation, albeit rarely.
Recent efforts to stochastically simulate gene expression have focused on reducing the number of states and complexity of the system being studied by approximating the kinetic delays that occur during transcriptional and translational elongation processes. For example, the stochastic simulator Dizzy allows for modeling transcription elongation as a series of equivalent steps that can be sampled in total from the gamma distribution, as Gibson and Bruck suggest
Thus, in order to efficiently and more realistically simulate base-pair resolved models of gene expression, we developed a single-molecule stochastic simulator, Tabasco, optimized to handle molecular events specific to gene expression such as the initiation, elongation and termination of transcription and translation as well as interactions among protein-DNA complexes. Tabasco tracks individual molecules and reactions on the DNA at single base resolution (e.g., RNA polymerase transcribing DNA) in order to avoid the specification of many unoccupied system states. Tabasco also uses dynamic reaction generation based on encoded rules for gene expression in order to avoid specification of all possible states and transitions within the system (e.g., all RNA polymerase molecules transcribe DNA to produce RNA). This framework provides the accurate descriptions of gene expression dynamics while allowing analysis of phenomena such as how intermolecular events between DNA-protein complexes affect system-wide gene expression.
Our motivations for creating Tabasco started from our interest in simulating the dynamics of bacteriophage T7 gene expression during phage infection. For example, past models of T7 infection used ODEs to simulate T7 gene expression dynamics. The use of ODEs limited our ability to accurately simulate the kinetics of T7 gene expression and study how intermolecular events may impact the genome-wide allocation of expression resources
Tabasco is a stochastic simulator that tracks individual molecules of DNA and associated proteins at single base-pair resolution. Tabasco makes use of a Gibson-accelerated Gillespie SSA to compute the reaction event timing and the resultant time-evolution of the genetic system
Tabasco transitions between two levels of resolution while simulating gene expression: "single-molecule" and "species level" (Figure
Figure 1
Structure of TABASCO simulator
Structure of TABASCO simulator. A simplified flow diagram of gene expression shows how Tabasco transitions from tracking individual molecules (Single Molecule Resolution) to grouping them into species (Species Level Resolution). The arrows represent reactions that can occur (process labeled in blue).
The transition between the single-molecule level and the species level occurs during the tracking of RNA abundances (Figure
Figure 2
Differences in single-step and multi-step elongation processes
Differences in single-step and multi-step elongation processes. Simulating the elongation of RNA polymerase along 50 based of DNA as a one-step elongation process versus a chain of individual elongation events gives the same average elongation time, but produces marked differences in the distribution of those elongation times. The process shown here is either a single 1.0 second elongation step (red) or fifty 0.02 second elongation steps (blue). Each histogram is the distribution of resulting times from 20,000 independent simulations.
The structure of Tabasco confers at least four advantages. First, treating gene expression at base-pair resolution allows for more accurate representation of the kinetics of gene expression. For example, traditional SSAs often lump multi-step reactions as single steps causing inaccurate estimates on pre-steady state kinetics. Second, tracking the state of individual proteins on DNA and allowing internal logic to automatically generate reactions eliminates the need to enumerate all the possible states of polymerases and proteins associated with the DNA. For example, transcribing polymerases and processes such as genome entry into a cell can cause certain protein binding sites to be inaccessible (Figure
Figure 3
Advantages of single molecule resolution
Advantages of single molecule resolution. Tracking individual molecules of DNA at a base-pair resolution allows for creation of implicit rules defined by the state of protein-DNA interactions and of the DNA itself. For example, when a traversing polymerase occupies a genetic element such as a protein-DNA binding site (a), the binding site is inactivated and prevented from binding a protein until the polymerase completely clears the site. Activation of genetic elements by entry of DNA into the cell (b) and inter-polymerase interactions (c) can be directly simulated as well. Finally, in complex genetic environments (d) where multiple promoters and terminators regulate the expression of genes, a priori transcription levels for each protein need not be calculated, for they are generated implicitly.
Simple gene expression models
As an initial test, we simulated the expression of a 1,000 amino acid protein using both Tabasco and a standard species-level SSA (Figure
Figure 4
Comparison of TABASCO to species-level simulations for a simple gene
Comparison of TABASCO to species-level simulations for a simple gene. We simulated the expression of a 1,000 amino acid protein using Tabasco (blue), and a standard species-level simulator (red) in which the processes of elongation (transcription and translation) were lumped into single steps. While the steady-state averages and standard deviations (a, b) are statistically equivalent, the pre-steady state kinetics (c) are quite different. Production of the first protein product (d) is much faster in the standard species-level simulator than in Tabasco. In fact, the species-level simulator generates unrealistic times to first production of a a protein; elongation of the polymerase alone should take ~75 seconds for a gene of this size (see text). The data shown are averages of 500 individual simulations.
Table 1
Constants used in the Simple Gene Expression Model
We compared two models of expression of a single-gene varying only if mRNA and protein elongation reactions are treated as series of individual steps or a single lumped reaction (Figure 3; Figure 6). The kclear rates do not exactly match because of small differences in the model structures (see text). The Tabasco constant for
The discrepancy in the time for synthesis of a first protein product occurs for two reasons
Polymerase Interactions
The first natural biological system that we studied using Tabasco is bacteriophage T7. During bacteriophage T7 infection, the
In order to test how different models of polymerase-polymerase interactions impact gene expression, we first used Tabasco to simulate gene expression from a reduced genetic system that captures key features from the layout of the T7 genome. Briefly, this two-gene system has an
Figure 5
Simulating models for inter-polymerase interactions
Simulating models for inter-polymerase interactions. Average steady-state protein levels from simulations of the DNA molecule with promoters for different polymerases (T7 and
Genome-scale models and simulation
In order to test the effectiveness of Tabasco on genome-scale systems, we simulated gene expression during bacteriophage T7 infection. The 39,937 base pair T7 genome, as represented in our test simulation, is comprised of 52 coding domains, 22 host and phage promoters, two transcriptional terminators, and three distinct transcriptional feedback loops; four T7 coding domains were not represented as they are the result of translational frameshifts or alternative start sites for other genes
Using realistic parameter sets, we simulated a single cell being simultaneously infected by three phage particles (Methods). The protein-DNA interactions on the DNA were tracked independently for each infecting phage, but use the same pools of soluble proteins in the cell; thus, these simulations track a combined 119,811 base pairs of DNA. We incorporated known mechanisms of cellular entry of T7 DNA into our simulations. We also developed a method to visualize the results of our simulations that allows for the graphical display of dynamic Tabasco output (Methods). Simulation of a full-scale genomic model of T7 gene expression on a single AMD Athlon MP 2100+ 1.8 GHz processor takes between 1–2 hours for a model that represents 30 minutes of real-time infection. Visualization of Tabasco output displays both T7 and
Additional File 1
Visualization of T7 genome simulation using Tabasco. We developed a general visualization tool for depicting graphically and dynamically depicting output from Tabasco. Here, we show output from a Tabasco simulation of three T7 phage simultaneously infecting a single cell over the course of 25 minutes. Each of the three white lines at the top represents a distinct copy of the 39,937 bp T7 genome. The T7 and
Click here for file
Discussion & Conclusion
We designed and developed Tabasco to revisit approximations encoded within previously available gene expression simulation algorithms, and to develop a method for efficiently computing single-molecule, base-pair resolved models of gene expression. Previous tools required either modeling smaller systems at increased resolution, supercomputers, or simplifications and assumptions of the biophysical models that result in reduced simulation accuracy. For example, Tabasco's implementation of a Gibson-accelerated Gillespie SSA allowed us to bypass simplifications that are often made for promoter binding by RNA polymerase and transcription initiation
Like Moleculizer, Tabasco dynamically generates reactions based on rules. Specifically, Tabasco uses the mechanics of gene expression as rules in order to avoid specification of all the states and transitions prior to simulation
Since Tabasco tracks individual molecules of DNA, the processing power required for Tabasco simulation scales with the copy number and length of the template DNA. While requiring significant processing power, Tabasco is computationally tractable in regimes for which stochastic simulations are often needed to produce accurate results (for example, when the number of DNA molecules in the system is low). Those interested in using simulators similar to Tabasco to study detailed interactions at the RNA level would face challenges due to the higher numbers of RNA compared to DNA in the cell. Such challenges may be overcome in the future by incorporating the work of others on increasing the efficiency of SSAs without significantly sacrificing accuracy
While Tabasco is an exact SSA, it only exactly simulates the kinetics of the already simplified models that we give it. For example, we model transcription elongation as a first-order reaction, even though we know that this process is more complicated
Executables, source code, documentation, and usage notes for Tabasco are freely available, and should facilitate future extensions on the current design (Availability and Requirements). We have already found Tabasco to be useful in constraining models of T7 gene expression by comparing simulator output to new experimental measurements of absolute copies of mRNA abundance during infection; these results will be presented in a forthcoming paper [Keller S, Endy D, Kosuri S,
Overall Simulator Structure
Tabasco implements a modified version of the Gibson Next Reaction Method (NRM)
The NRM uses a dependency graph to determine which reactions are affected by any particular reaction's execution. In the NRM, the dependency graph is constructed only once at the start of simulation and remains unchanged afterwards. However, since Tabasco creates reactions and complexes at the single-molecule level dynamically during simulation, a static dependency graph and indexed priority queue will not work. In order to solve this problem, Tabasco contains two specialized classes per DNA molecule within the overall indexed priority queue that are used to track a set of dynamically generated reactions. Each class contains a dynamic priority queue that stores the dynamically generated transcriptional and translational reactions and their tentative reaction times, as well the dependencies of any particular reaction (these specialized priority queues and the overall indexed priority queue are easy to confuse, and thus we will refer to the prior as the dynamic priority queue). The minimum tentative reaction time for all the dynamic reactions is set as the tentative reaction time of that dynamic priority queue with respect to the overall priority queue. Since, as in the NRM, Tabasco uses absolute times for determining the next reaction, the particular choice of the data structures containing the reactions does not affect the simulation results. The reason to separate the dynamic queues from the main indexed priority queue is to allow the size of the dynamic queues to change over time. As long as all dependencies are accurately updated upon reaction execution in the dynamic priority queues, the results are equivalent to the NRM. The structure of these dynamic priority queues will now be discussed in more detail.
A special class tracks the transcriptional processes for each DNA molecule. This class contains a dynamic priority queue that keeps track of all reactions related to that DNA molecule, such as transcriptional elongation processes. All reactions that are stored in this dynamic priority queue have tentative reaction times (as calculated by the NRM), and the minimum tentative reaction time is used to set when this class should be called to execute within the indexed priority queue. In order to determine what the effects of a particular binding or elongation reaction are, the class contains two arrays where each element represents one DNA base. Each element of the first array contains pointers to transcriptional elements encoded at that location such as promoters and terminators. The second array contains pointers to all DNA-protein complexes that reside on the DNA such as elongating RNA polymerase. Only one transcriptional control element or DNA complex can occupy any particular position at any time.
The framework we use here to simulate protein-DNA binding is shown in Figure
Figure 6
Model structures for Tabasco and species-level simulations of expression of a single gene
Model structures for Tabasco and species-level simulations of expression of a single gene. We simulated expression of a single gene using a standard species-level SSA, and using the Tabasco gene expression simulator (Figure 3). The Tabasco-based simulation uses a transcription initiation model (a) with the rest of gene expression using the general schematic shown in Figure 1. We constructed the model for the species-level simulator (b) to mimic the Tabasco-based model, except that elongation reactions were treated as single-lumped reactions.
Once an elongation complex is formed, each elongation reaction causes the complex to move one base pair, and the elongation reaction is updated with a new time for the next elongation step. When an elongation reaction executes, the algorithm also checks whether another complex's footprint prevents the current polymerase from moving forward by checking the array containing all complex positions on the DNA. If there is another polymerase blocking the current polymerase's path, the polymerases will behave according to the polymerase interaction model chosen at the start of simulation. For example, the upstream polymerase can terminate, or signal the downstream polymerase to terminate the next time the polymerase is set to elongate, or simply hold its position until the next opportunity to elongate. The elongation reaction also does a check for transcriptional elements on the DNA such as promoters and transcriptional terminators. Upon arriving at a transcriptional terminator, a polymerase will either continue transcribing or terminate depending on a stochastic decision that will also be discussed below. If a transcribing polymerase occludes an open promoter, then the promoter will be decremented and unavailable for binding.
The "stochastic decisions" described above for transcriptional termination and abortive initiation on the promoter are used as methods to simplify parameterization in the models, so as to be consistent with how experimental data for such events is typically reported. In the case of termination, a uniform random number is chosen, and if that number is less than the termination efficiency, the polymerase will terminate transcription and fall off the DNA. Analogously, in the case of abortive initiation, if the uniform random number is less than the abortive initiation percentage, then the next reaction will be an abortive initiation step rather than formation of an elongation complex. These decisions take no simulator time, but cause changes in the downstream actions the complexes may take. This simplification is not done for computational efficiency, but more for model simplicity. Tabasco could be modified, with little computational load, to replace the stochastic decisions into competing rates of different reactions. However, these rates are not well understood, and thus we chose to incorporate the measurable models of termination efficiencies and abortive initiation frequencies into the simulator.
The transcribing RNA polymerase complexes also produce mRNA, which are tracked by a separate class. Since there are many more copies of RNA than DNA during simulation, and because we were uncertain as to the importance of protein-protein interactions on the mRNA, we chose to treat the majority of translation at the species level. However, if, for example, an RNA polymerase prematurely terminates before reaching the translation stop site, the mRNA and the ribosomes translating it will not produce function proteins. Thus, so long as the coding sequence is still being transcribed, we must treat all translation events at the single-molecule level as well. However, as soon as the entire coding sequence of an open reading frame is transcribed, Tabasco transitions to tracking species of mRNA molecules.
We used two classes to model the formation of RBSs and their respective start sites, the Nascent RBSs and Mature RBSs, in order to differentiate when mRNA should be tracked at the single-molecule level or the species-level, respectively. At the start of simulation, one more array of genome length is created that contains the positions of translation start and stop sites on the DNA. As RNA polymerase elongates (as described above), if the polymerase transcribes past a translational start site, a Nascent RBS is made. This Nascent RBS is available for binding by free ribosomes, and a reaction is automatically created and placed into the dynamic priority queue for translation processes. Once the polymerase arrives at the corresponding translation termination site, the Nascent RBS is converted to a Mature RBS, which is tracked at the species-level in the overall Indexed Priority Queue.
Translation, both at the single-molecule level and species level, occurs in three steps. First, ribosomes can bind either Nascent or Mature RBSs; there is an initial reaction to form the (Nascent or Mature) Initiation Complex. Second, these initiation complexes are converted to (Nascent or Mature) Elongation Complexes and an RBS at a rate that depends on the speed of the ribosome and the length the ribosome must travel to clear the ribosome binding site. This reaction is treated as a single step, however the distribution of times is chosen from a gamma distribution, in order to better represent a series of individual elongation steps. Third, the Elongation Complex then forms a finished protein and a free ribosome at a rate proportional to the remaining length of the open reading frame. This reaction also is computed via a gamma distribution.
At the single molecule level, two additional mechanisms allow for the state of the transcribing RNA polymerase to affect translations. First, if the transcribing RNA polymerase terminates before reaching a corresponding translation stop site, all bound ribosomes are immediately released, the Nascent RBS is removed, and no protein product is formed. Also, for any particular Nascent RNA, the maximal number of Nascent Elongation Complexes that can be formed is capped at the length of the currently transcribed portion of the open reading frame divided by the footprint of the ribosome.
Finally, to note, all transitions between single-molecule tracking and species-level tracking occur when a reaction, such as RNA polymerase termination or Initiation Complex conversion into an Elongation Complex. Thus, the transition from single-molecule tracking back to the species level tracking should also not affect the validity of using the NRM.
DNA entry
We developed multiple models to represent DNA entry into a cell or compartment (such a step can be useful in starting a simulation of infection or transformation). First, DNA can enter the cell via a zero-order constant reaction rate. Second, RNA polymerases have themselves been implicated as molecular motors that can drive DNA entry
Simulation, Data Output, Visualization, Code
Tabasco is written in Java® 1.4. The input file to the simulator is an XML file that describes and parameterizes the relevant genetic elements, initial conditions, and any other reactions that occur. The visualization is created by producing images that are then merged using Quicktime® to create a movie. The source code, executables, along with documentation and instructions for use are available within Additional Files
Additional File 2
Input files for simulations described in the text of the paper.
Click here for file
Additional File 3
TABASCO website.
Click here for file
Additional File 4
Parameter definitions within the input file. The table describes the parameter definitions that are used in the input files that are contained in the supplementary materials.
Click here for file
All constants used are provided for completeness. Please see the Supplementary Materials for input files and exact constants used.
Gamma versus Exponential distribution
The distribution of expected times for a reaction to occur in a stochastic simulator depends on the underlying model (Figure
Simple Gene Expression model
We simulated two models of a promoter driving expression of a coding domain. The first model, termed single-molecule simulation, used the described Tabasco simulator to account for each reaction step during transcription and translation. The model uses the schemes shown in Figures
Polymerase Interactions & Bacteriophage T7
The simulations for the polymerase interactions and bacteriophage T7 development used parameters that can be found in the input files in the Supplementary Materials. Table S1 details the meaning of each of the parameters in the input files. The parameters used were based on empirical measurements where possible. However, in general, the exact values of the constants are ancillary to this section, which is to show that Tabasco is able to simulate processes such as polymerase interactions and entire genetic systems such as T7. The parameter derivations are detailed are detailed elsewhere
Availability and Requirements
Project name: TABASCO;
Project home page:
Operating system(s): Platform independent;
Programming language: Java;
Other requirements: J2SE 1.4.2 or higher
License: Public domain;
Any restrictions to use by non-academics: n/a.
Authors' contributions
SK and DE developed the idea of Tabasco. SK did all work on Tabasco design, code development, modeling and simulation, and drafted the manuscript. SK and JRK developed the Tabasco visualization software. SK and DE edited the manuscript. All authors read and approved the final manuscript.
We thank Mark Allen Weiss and Jonathan Goler for use of source code for the priority queue implementation and XML parser, respectively. We thank Kirsten Benjamin for providing pre-publication results of the number of Ste5 molecules in a yeast cell. We thank Heather Keller, Ty Thomson, and Rebecca Ward for critical reading of earlier versions of this manuscript. This research was supported by grants to DE from the US NSF, DARPA, and NIH. SK received additional support via an Andrew and Edna Viterbi Fellowship in Computational Biology and the MIT/NIGMS Biotechnology Training Program Fellowship. Additional support was provided by MIT.
Single-molecule analysis of RNA polymerase transcription
Promoter clearance and escape in prokaryotes
Initial transcription by RNA polymerase proceeds through a DNA-scrunching mechanism
Abortive initiation and productive initiation by RNA polymerase involve DNA scrunching
The meaning of systems biology
Stochastic mechanisms in gene expression
Optimal design of feedback control by inhibition
Optimal design of feedback control by inhibition: dynamic considerations
Stochastic simulation of chemical kinetics
Deviant effects in molecular reaction pathways
Predicting temporal fluctuations in an intracellular signalling pathway
Quantitative functional interrelations within the cis-regulatory system of the S. purpuratus Endo16 gene
Cis-regulatory logic in the endo16 gene: switching from a specification to a differentiation mode of control
Stochastic simulation of chemical reactions with spatial resolution and single molecule detail
Automatic generation of cellular reaction networks with Moleculizer 1.0
Dizzy: stochastic simulation of large-scale genetic regulatory networks
Efficient exact stochastic simulation of chemical systems with many species and many channels
Validation of an algorithm for delay stochastic simulation of transcription and translation in prokaryotic gene expression
Stochastic kinetics description of a simple transcription model
Intracellular kinetics of a growing virus: A genetically structured simulation for bacteriophage T7
Computation, prediction, and experimental tests of fitness for bacteriophage T7 mutants with permuted genomes
Delay-induced stochastic oscillations in gene regulation
Computer simulation of T3/T7 phage infection using lag times
Gene expression time delays and Turing pattern formation systems
Autoinhibition with transcriptional delay: a simple mechanism for the zebrafish somitogenesis oscillator
Oscillatory expression of Hes1, p53, and NF-kappaB driven by transcriptional time delays
The use of delay differential equations in chemical kinetics
Organization and expression of bacteriophage T7 DNA
Rate of translocation of bacteriophage T7 DNA across the membranes of Escherichia coli
Promoter occlusion: transcription through a promoter may inhibit its activity
Cooperation between RNA polymerase molecules in transcription elongation
Regulation of transcription from tandem and convergent promoters
Observed instability of T7 RNA polymerase elongation complexes can be dominated by collision-induced "bumping"
Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements
The OR control system ofbacteriophage lambda. A physical-chemical model for gene regulation
Multiscale stochastic simulation algorithm with stochastic partial equilibrium assumption for chemically reacting systems
Efficient step size selection for the tau-leaping simulation method
Improved leap-size selection for accelerated stochastic simulation
Stochastic chemical kinetics and the quasi-steady-state assumption: Application to the Gillespie algorithm
An integrated model of the transcription complex in elongation, termination, and editing
Probing transcription factor dynamics at the single-molecule level in a living cell
Facilitated target location in biological systems
RNA polymerase-dependent mechanism for the stepwise T7 phage DNA transport from the virion into E. coli
Numerical recipes in C : the art of scientific computing
The art of computer programming
Simulation, Models, and Refactoring of Bacteriophage T7


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The Western Conference is as loaded as Tony La Russa behind the wheel of a moving vehicle. 10 team post winning records, with 9 of them having 33+ wins as of February 21st. Only 5 games separate the team in 1st (New Orleans) and the team in 9th (Golden State). If the season ended today, the Warriors would miss the playoffs, but if they were in the Eastern Conference, the Bay Area would be hosting a first round match up. The West was already wild, but all these trades have made it impossible to pick a favorite.
Shaq and Jason Kidd are both 0-1 in their debuts from last night. It's going to take a little time for both of them to adjust. Shaq seems like a square piece in a round hole. He did play 29 minutes in just his first game in Phoenix, scoring 15 and pulling down 9 rebounds. Not a bad first showing, but they did allow 130 points and lose. Kidd had only 5 assists and 8 points for Dallas, as Chris Paul straight owned him.
Here are the 10 teams above .500 in the West, core players, with new acquisitions in bold. (In order of current standings.)
New Orleans Hornets:
Chris Paul, David West, Tyson Chandler, Peja Stojakovic, ( Mike James, Bonzi Wells, if trade goes through)
Chris Paul is my MVP as of right now. Last night, he had 31 points, 11 assists, and 9 steals against J-Kidd and the Mavs. He's averaging 21-11 and shooting a pretty good percentage. West and Chandler can do all the dirty work and Peja is back shooting from deep. I'm not sure they can hold on to the top spot in the West or win the whole thing, but certainly not a team you would want to face. Can Paul carry them to the Finals?
Los Angeles Lakers:
Kobe Bryant, Pau Gasol, Lamar Odom, Andrew Bynum, Derek Fisher, and the bench mob
Right now I'm leaning towards picking the Lakers to win the West, even though I'm a Celtics fan and hate the Lakers. The addition of Gasol, or should I say the thievery that was the Gasol deal, has put this team over the top. The Lakers are 7-1 with Pau. It's going to be interesting to see how he and Bynum play together, but suddenly they have Odom playing a role that he is much more comfortable with. They don't need to count on him to score, but he still does. Kobe is Kobe, and these guys look like they are headed to the NBA Finals.
Phoenix Suns:
Steve Nash, Amare Stoudemire, Shaquille O'Neal, Grant Hill, Leandro Barbosa, Raja Bell, Boris Diaw
You can't get crazy about last night no matter what happened in Shaq's debut. It's going to take more than 1 game to see how the deal will play out. Amare is going to put up ridiculous numbers (37-15 last night), but it does not matter if they lose 130s to 120s. Who's playing defense on this team? Raja Bell? He gave up 40 last night to Kobe. Nash, Amare, and Shaq at this stage of his career, are not going to scare you at the defense end. I'm looking forward to Friday when Boston comes to town. Shaq's first 3 games in Phoenix: Lakers, [[Boston Celtics|Celtics[[, Pistons. They could go 0-3. When they sent Shawn Marion to Miami, they were the number 1 seed in the West. Now I don't think they host a 1st round series. I know they had not made the Finals with the current Big 3 of their own (Nash, Amare, Matrix), but this year with the make up of the West I thought they could have. We'll see.
Utah Jazz:
Deron Williams, Carlos Boozer, Andrei Kirilenko, Mehmet Okur, Kyle Korver, Ronnie Brewer, Paul Millsap, Matt Harping
It doesn't get a whole better for a 1-2 punch in basketball than Williams and Boozer. The addition of Korver has given the Jazz a whole new element to their attack. I love the combination of talents they have here, with bangers like Boozer and Millsap, shooters in Okur and Korver, slashers in AK-47 and Brewer, and of course one of the top point guards in the game in Williams. This is just another team in the West that you could not be shocked if they won it all. Early prediction is that the Jazz lose in the conference finals to the Lakers.
San Antonio Spurs:
Tim Duncan, Tony Parker, Manu Ginobili, Bruce Bowen, Michael Finley, Damon Stoudamire, Kurt Thomas.
No one is talking about the Spurs. Maybe we should be. As soon as you count them out, you'll see Tim Duncan holding the Championship trophy in the air for 5th time in his career. But still, like everyone else, I'm not convinced they will be there. They have been winning it every other year, so maybe it's not meant to be. What a great gift for a top 4 seed it would be to have to face the Spurs. Good luck.
Dallas Mavericks:
Dirk Nowitzki, Jason Kidd, Josh Howard, Jason Terry, Jerry Stackhouse, Brandon Bass, Eric Dampier.
I'll be honest: I liked the Kidd deal, and I don't think that they gave up too much to get them. Devin Harris and what else? The two first round picks will be low and a guy like Antoine Wright is only 24 and will be better than anyone they could get at the end of this years draft. Kidd still has something left in the tank, his rebounding and assists numbers are huge this year even though he has struggled shooting from the field. I did not think they could win the whole thing prior to the Kidd trade, now they at least have a chance.
Houston Rockets:
Yao Ming, Tracy McGrady, Shane Battier, Skip to my Lou, Luis Scola, Bobby Jackson (if trade goes), Luther Head, Carl Landry
I think the Rockets end up missing the playoffs. Even if they get in you can't like T-Mac's chances of advancing. It looks like they might be parting with Mike James and Bonzi Wells, and bringing in veteran point guard Bobby Jackson. (Jackson would be great in a Celtic's uniform.) Despite being just 4.5 games out of first the Rockets will be fighting to make the playoffs for the rest of the year.
Denver Nuggets:
Allen Iverson, Carmelo Anthony, Marcus Camby, Kenyon Martin, Anthony Carter, J.R. Smith, Linas Kleiza.
Is there a more talented starting lineup in the league? Maybe not. But they have always had a hard time putting it all together. I think they do get in the playoffs even though they are one of the only teams in the West not to make a move. (As of now). These guys can obviously beat anyone on any given night, but what about 4 out of 7? You know they will score enough points. The question remains, is Camby enough on defense?
Golden State Warriors:
Baron Davis, Stephen Jackson, Monta Ellis, Al Harrington, Andris Biedrins, Chris Webber, Matt Barnes, Keleena Azubuike, Mickael Pietrus.
The Chris Webber addition doesn't look like it's going to be much of a boost. He starts sure; I don't see him as a major factor however. The way they shoot the 3, early and often, they can certainly beat anyone. Like the Nuggets it will be difficult for the Warriors to advance far in the playoffs. I know they beat the Mavs last year in the 1-8 match up. I say they do get in the playoffs led by Baron Davis, but it ends in the 1 st round this year.
Portland Trail Blazers:
Brandon Roy, LaMarcus Aldrige, Travis Outlaw, Jarrett Jack, Martell Webster, Steve Blake
It was a nice run early in the season; they just could not keep up with all these great teams in the West. At 28-25, they have still surpassed expectations without Greg Oden. Definitely keep an eye on this team going forward. Brandon Roy has been great to watch and they play team basketball. Of the 10 teams on this list, I have the Blazers and Rockets missing out on the playoffs in the end.
This list could get a shake up as the NBA trade deadline is minutes away. Players like Vince Carter, Zach Randolph, Ron Artest, Ben Wallace, and more could be moving around. Who else will make a move? The next couple of months will be great, a lot of questions still left to be answered.
-Rich Keefe
Check out all our stuff at http://sportsbrief.blogspot.com/ and e-mail us at firstname.lastname@example.com


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Gaia Royal Hotel is a very large holiday resort, set on the beach, with a real village style feel, with its main building and several two-floor bungalows, offering fantastic holidays. In the main building is the original hotel lobby and reception, lounge with satellite TV area, bar, restaurant with terrace and a mini market. Outside, all set in mature large gardens are 3 large freshwater swimming pools with 3 separate children pools, deckchairs and umbrellas as well as two pool-snack bars.
Gaia Royal Hotel is situated at the holiday resort Mastichari, only 100m from the very long sandy beach away and about 5km from the fishing village Mastichari, where many taverns, bars and shops are situated. A regular bus stop within 500m with schedule to Kos Town . Sun chairs and umbrellas at the beach are at a local charge.
The comfortable and very tastefully decorated rooms have bath or shower/WC, hairdryer, radio, telephone, air conditioning, balcony or terrace. The hotel offers 3 pools surrounded by sun terraces with sun beds and parasols and the pool bars are ready to serve you cooling drinks all day. For children you'll find kids pools, a kids playground area, and a kids club- however- English is not spoken by the kids club staff.
* 4 Bedded Rooms
* Beachfront
* Children's playground
* Children's pool
* Lounge
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* Reception - 24 hrs
* Sunbeds


text: Tips for off-season sports bettors By: SBR | Tuesday, February 3, 2015 With the Super Bowl out of the way, there has never been a more important time to make sure you are playing with a well-funded and reputable online sportsbook. Many bookmakers deal with their biggest rush of withdrawals from North American players during this time-frame, so the importance of sticking with a proven establishment cannot be understated. The following tips should help guide you in the right direction and keep your bankroll as safe as possible. #1: Use sportsbook rating information The letter grade system in the sportsbook ratings guide was not thought of overnight. Some of the sportsbooks listed in the comprehensive guide have been online since the late '90s. Ratings are for not sale and have been earned by merit. #2: Consider what playes like you have to say The annual SBR Posters Poll is usually a good indication of what's shaking in the industry. To earn a spot in the top five is no small task, and the bookmakers selected by a community of veteran posters should not be slept on. #3: Manage your bankroll With so many free handicapping resources at your disposal, there should be no compelling reason to avoid using tools such as the half-point calculator. Every small edge counts, and picking the right online sportsbook is only half the battle. Staying in the game is the other. If you find yourself in a dispute with an online sportsbook, all hope is not list. Submit a sportsbook complaint with SBR and an analyst will review your ticket free of charge and advise you on what happens next.


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There are certain moments as a parent you wish you could just bottle up and save forever.
I am grateful for an acute awareness of how short "these days" of young childhood are. Like that feeling of your four year old's small hand resting on yours, their head leaning on your shoulder and the warmth of their little body as they stretch out next to you. It's a scene that plays out again and again in our house as we greet the night with story time. No matter how good/bad/crazy life has been, this is how we close the day. It is, perhaps, one of the most cherished things I do get to do each day with the boys.
Reading is the gateway to all opportunity in our world. Much of parenting culture these days is fixated on giving kids an "edge." It can sometimes feel like reading has become another hoop for kids to jump through - the earlier they can do it on their own the better. There literally are flash card systems for infants to start recognizing sight words. All of this makes me cringe. If there's anything I want to teach my boys about reading right now, it's that it is fun, warm, cozy and interesting. Who better to teach them than Mom or Dad? Most children are actually not ready to start learning to read in earnest until somewhere between ages six and seven. Studies show that earlier attempts to teach reading backfire and result in reduced interest in reading. This makes sense, who enjoys being asked to do something over and over again that you simply cannot do?
In contrast, being read to is something that every child, even infants, really enjoy. Why? Well, first of all, I challenge you to try to read a book to a small child and not cuddle them! It's next to impossible. They inevitably end up on your lap, leaning against you, or at least sitting close. The resultant shot of vitamin L (AKA love) is potent. They get to hear the voice of their favorite person (you!), saying a variety of interesting words, and for the most part they get to look at interesting pictures. You can take your reading to the next level by pointing at interesting things in the book and talking about what you see. All of this exposes children to a variety of new and learned words, which has been shown to improve literacy later in life. This association is so well recognized that this week, the American Academy of Pediatrics recommended that parents read to their infants on a daily basis.
There are books on our shelves that I have probably read hundreds of times ("In the great green room…"). Every parent can attest to kid's love of reading the same books, over and over (and over) again. It can seem tedious; we are stuck on a particular National Geographic photo book about the Indian Ocean right now, but we really try to stick with it. Why? Science tells us that young children really enjoy knowing what comes next. Repetition allows them to lay down long lasting and enduring neural pathways and allows them to feel as though they have "mastered" a skill. My toddler loves it when I pause and allow him to finish a sentence in his favorite book.
When you read to your child, you are laying down foundations. A love of reading, a habit of spending time together, a tradition that can be passed on when your babies grow up and begin reading to their own children. There are books on our shelf that leave me with a frog in my throat, Love you Forever by Robert Munsch, for example. It's the same story that my parents read to me when I was a little girl. The story is about a mother's enduring love for her son with the passing of life's seasons, and I feel the poignancy of that passage of time in my own life with every turn of the page. It is beautiful and inspiring, and as I read the last verse of the story with my little boys curled up next to me, it feels like coming home every time.
Jan Sommers is a local physician. She lives with her two young sons and husband Ryan Sommers in Truro.


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Print Artist Platinum 25 is all-rounder software for making of print with more than 16000 artistically designed templates and mesmerizing models.
This versatile software contains various useful features that are given below:
- Robust text tools.
- Photo editor for cropping, trimming etc.
- Function for Red-eye removal.
- Repairing of low-lighting photos.
- Compatible with all operating systems of Windows.
- User-friendly interface.
User Rating ( votes)


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Chef Manuel Perez will create a menu for your event that is truly representative of your tastes, style, budget, and theme. His "New American" cuisine is receiving rave reviews (voted top 25 restaurants in New Jersey) and his dishes will no doubt both tantalize and impress your guests. The Peacock Inn can also work with you on recommendations for Princeton's finest flowers, entertainment, decorations, audio/visual equipment, and more.
Bed and Breakfast & Inns Boutique Hotels Hotels Small Luxury Hotels Unique Event Locations


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Incorreggibili NinfomaniFavorites [+]
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DRM-free downloads offer content that is not limited by standard Digital Rights Management (DRM) access technologies. These technologies allow hardware manufacturers, publishers, copyright holders and individuals to limit the usage of digital content.
When you purchase DRM-free downloadable content, that content is available for you to download on any computer and save to a disk whenever you like. You can also download to multiple computers. There is no timeframe associated with your purchase and the content will not expire.
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Youtube is more than entertainment and can provide schools an efficient, cost effective and pratical method for curating, sharing and editing video. This session will focus on how teachers can set up a channel to currate and assess student created projects. With the help of an iPad or iPod any student can create projects, and upload their work and edit their projects on youtube. Privacy and safety issues often prevent teachers from taking advantage of youtube and this session will discuss methods for safely using Youtube in school.
Presenter: Brian Casey
2012 Archive > 2012 Sessions >


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How to Have a simple conversation in Hungarian
Check out this instructional Hungarian language video to learn how to speak Hungarian! In the 1st lesson, listen to tiny dialogues in Hungarian. This video provides conversations exclusively in Hungarian without English translation or subtitles. This instructional language video is better for advanced Hungarian speakers who need to improve their listening skills.


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Leverage Bing Custom Search to build a search that fits your needs with powerful ranking, a global-scale search index, and document processing. Setup is easy and fast. The core technology works in three steps: it identifies on-topic sites, applies the Bing ranker, and delivers relevant search results while allowing you to adjust the parameters at any time.
Customize your search
Quickly and reliably define the slices of the web that you want to draw from. Change the parameters of the sites you want and don't want at any time. Explore site suggestions to intelligently expand the scope of your search domain.
Custom Search features a straightforward UI that enables you to create your web search without a line of code.
Usage is ad-free – no matter how much or how little of the service you use.
Learn more
Check out the other Cognitive Services APIs
Computer Vision API
Distill actionable information from images
Content Moderator
Automated image, text, and video moderation
Video API PREVIEW
Intelligent video processing
Video Indexer PREVIEW
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Face API
Detect, analyze, organize, and tag faces in photos
Emotion API PREVIEW
Personalize user experiences with emotion recognition
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Teach your apps to understand commands from your users
Bing Spell Check API
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Enrich your experiences by identifying and augmenting entity information from the web.
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text:
Since Epic EEA Announcements 10 has finally come, Red Inc. can start making lots of more games again. But, it is currently unknown if Red Inc. will start making Super Mario Adventures II. Keep watch on the Red Inc. News to see if this project will be started or abondoned. If we abandon it, any user is free to adopt it and make it theirself.
Red Inc. also announced the two Mario Jet Blast and RedYoshi DS RPG games today. We'll be working on MJB alone, but Bomb Productions Games will help out with RY: DS RPG.


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Home > Product Categories > Batteries and Power > Batteries & Chargers > Camera Battery Mounts > QR-A200
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Adapts the wedge mount for the Sony DNWA225/DNWA220/DNWA25DSR70 portable SX editing machine to the Anton/Bauer Gold Mount. Because of the high current be used to power Sony DSR 50VTR and BVMD9 High Definition Monitor.
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text: LTE and smartphone uptake drives video traffic growth says Ericsson07 Jun 2013 10:08 AM Ericsson's latest Mobility Report indicates that video traffic growing by 60% annually, driven by better network speeds, while total global smartphone subscriptions hit the 1.2 billion mark in 2012The new edition of the Ericsson Mobility Report reveals that mobile-data traffic will continue to grow significantly in the coming years, a trend driven mainly by video. Overall data traffic is expected to grow 12-fold by the end of 2018.Increasing usage is driven by continual growth in the amount of content available as well as the improved network speeds that come with HSPA and LTE development.Douglas Gilstrap, Senior Vice President and Head of Strategy at Ericsson, said: 'LTE services will be available to about 60% of the world's population in 2018. We expect LTE subscriptions to exceed 1 billion in 2017, driven by more capable devices and demand for data-intensive services such as video. Owing to the build out of WCDMA/HSPA, network speeds have improved, and so has the user experience.'Video makes up the largest segment of data traffic in networks, and it is expected to grow around 60% annually up until the end of 2018. Video consumption is on average 2.6GB per subscription per month in some networks.While video is popular, users don't necessarily tend to spend the most time on data-heavy applications. Consumers spend more time on social networking: an average of up to 85 minutes per day in some networks.Smartphones accounted for around half of all mobile-phone sales in Q1 2013, compared with roughly 40% for the whole of 2012. The number of total mobile subscriptions grew by 8% globally year-on-year by Q1 2013.Of those, WCDMA/HSPA added around 60 million subscriptions, GSM/EDGE-only subscriptions grew by roughly 30 million, and LTE added around 20 million new subscriptions. Mobile-broadband subscriptions grew even faster over this period (at a rate of 45% year-on-year), reaching around 1.7 billion.The Mobility Report also addresses the concept of 'app coverage' - broadening the definition of coverage from voice to include how well users are able to access their mobile apps - and presents a new framework explaining the effects of varying network performance in a way that is relevant to the user.In this edition of the report, the relationship between network performance and consumer loyalty was also studied. Network performance is the principal driver of subscriber loyalty to mobile operators, followed by value for money. Other sections of the report explain the effects of smartphone signalling on data traffic, and look at data roaming, identifying opportunities for operators to generate new revenue streams.Wireless magazineLeave a CommentPlease write code to prove you're humanMore on... LTE GeneralMWC17: Zyxel to debut new LTE and Wi-Fi indoor gatewayHuawei, Vodafone and Qualcomm create LAA ready networkNokia demos live private LTE network over CBRS shared spectrumMore on... EricssonMWC17: Ericsson expands 5G access and transport portfolioOperators trial intercontinental 5G roaming using network slicing Latest in LTEMWC17: Zyxel to debut new LTE and Wi-Fi indoor gatewayHuawei, Vodafone and Qualcomm create LAA ready networkNokia demos live private LTE network over CBRS shared spectrum


text:
By Tim Hepher and Matthias Blamont, Reuters
TOULOUSE, France European planemaker Airbus signaled the start of a downward cycle in production as global recession curbs jetliner demand, saying deliveries could fall and orders were set to tumble in 2009.
Airbus sales chief John Leahy literally rubbed a crystal ball in front of 200 journalists to highlight the difficulty of making predictions amidst the economic crisis, but said Airbus would probably sell at the lower end of 300-400 planes in 2009.
"It will be a very soft year," Leahy told a news conference on Thursday.
Airbus held the top spot in global airliner production for the sixth year running in 2008 with a 7% rise in reported deliveries to a record 483 planes.
Airbus also confirmed it had outsold Boeing last year with net orders for 777 planes, down 42% from 2007, though figures from both planemakers show the economy halting a three-year aviation boom.
Both Airbus and rival Boeing are bracing for more turbulence in an industry damaged by recent fuel price spikes as the economic downturn hits air travel. They also face a battle to prevent airlines canceling or deferring old orders.
Shares in parent company EADS rose 0.9%.
Airbus sold planes worth $100 billion in 2008, giving it a market share of 54% and lifting its backlog to 3,715 jets — just one more than rival Boeing, whose sales had been expected to cool after it smashed an industry record in 2007.
Asked if Airbus was profitable following two years of losses, Chief Executive Tom Enders said: "I would say so, yes."
EADS will publish results on March 10.
Boeing's 2008 production was hit by a 58-day machinists' strike with deliveries falling 15% to 375 planes. Boeing sold 662 aircraft, down 53%, it said last week.
The latest figures mean global large jetliner production shared by the two companies fell 4% to 858 planes last year. But analysts say steeper falls may be necessary as airlines cancel or defer orders to cope with the financial rout.
Airbus's prediction that it will sell fewer planes than it makes in 2009 means it will be the first year either big jetmaker has failed to replenish its backlog since 2003.
Enders said while he expected the level of deliveries to be in the "same ballpark" as the 2008 record, Airbus had drawn up contingency plans to cut output if necessary.
"It is more difficult than ever to predict the order intake, but I estimate the book-to-bill (order to delivery) ratio will be below one in 2009," he said.
Superjumbo orders
Airbus cut its A380 superjumbo delivery forecast for 2009 to 18 planes after recently warning it would miss its previous target of 21 by a "couple" of planes. Sales chief John Leahy said Airbus expected to sell 10 superjumbos in 2009.
Airbus said Air Austral, of the French overseas department of La Reunion, had provisionally bought two A380s in a single-class configuration, meaning it could seat over 800.
Airbus's biggest concern for 2009 is to ensure it does not end up making planes which it cannot sell as airlines starved of credit renege on past contracts at the last minute.
"The focus this year needs to be protecting deliveries," Enders said. "We are examining the situation on a daily basis."
To prevent unwanted planes known as "white tails" after the missing airline livery filling their tarmacs, planemakers can offer credit, line up alternative buyers or cut production.
Airbus recently sold jets discarded by India's Kingfisher to Nigerian carrier Arik, but such replacement deals are rare.
EADS said on Tuesday it was conserving cash and freezing acquisitions to help prop up aircraft sales by offering credit. Airbus financing was at a 20-year low above $1 billion in 2008 but it says it could absorb up to $5-6 billion.
"Both traffic and financing are an issue. Financing is a more immediate issue but both are essential to the outlook," Natixis Securities analyst Olivier Brochet said.
Airlines group IATA has said the industry was "shrinking by all measures" after a 4.6% traffic drop in November.
Boeing last week announced 4,500 job cuts.
Additional reporting by Kerstin Doerr.
Copyright 2009 Reuters Limited.
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US: Ford cuts truck production in US
Ford plans to cut make further cuts to pickup and SUV production in the US as the credit crunch bites.
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Senator John McCain fiercely defends his immigration proposals to angry suburban Arizona crowd
- Arizona crowd shouted down McCain's immigration proposals, saying immigrants are 'illiterate' and should be kept out of the U.S.
- McCain said there are too many immigrants living here illegally and 'we are not going to get enough buses to deport them'
- Homeland Security Secretary Napolitano was also visiting Arizona to discuss immigration reform Tuesday
- Visits from high-profile officials signify that Arizona will take center stage in national immigration debate
Sen. John McCain grew agitated Tuesday as he tried to defend his immigration plan before an angry crowd in suburban Phoenix who charged that reforms were moving too slowly.
'There are 11 million people living here illegally,' McCain, R-Ariz., said in response to members of the crowd who urged mass deportation of illegal immigrants. 'We are not going to get enough buses to deport them.'
McCain accused one man in the audience of not 'telling the truth' about illegal immigrants and added, 'They mow our lawns, they care for our babies, they clean, they wash up. That's what those people do, sir.'
Scroll down for video
Arizona residents became outraged at a town hall forum on immigration with U.S. Sen. John McCain, R-Arizona (right)
McCain defended his proposed immigration overhaul to an angry crowd in suburban Arizona in the latest sign that this border state will play a prominent role in the national immigration reform debate
Arizona resident Keith Smith shouted back at McCain, 'Cut off their welfare and all their stuff and they'll go back!'
'You said build the dang fence!' Smith added. 'Where's the fence?'
McCain turned his back to the crowd and pointed at a graph and the man shouted at him, 'That's not a fence!'
McCain snapped back: 'It's a banana. We've put up a banana without about $600 million dollars worth of appropriations.'
McCain said immigration reform should be contingent on better border security that must rely largely on technology able to detect border crossings. He said a tamper-proof Social Security card would help combat identity fraud, and noted any path to citizenship must require immigrants to learn English, cover back taxes and pay fines for breaking immigration laws.
Audience members shouted out their disapproval.
Audience members shouted out their disapproval. One man yelled that only guns would discourage illegal immigration while another complained that illegal immigrants should never be able to become citizens
One man yelled that only guns would discourage illegal immigration. Another man complained that illegal immigrants should never be able to become citizens or vote. A third man said illegal immigrants were illiterate invaders who wanted free government benefits.
'This is an overwhelming experience,' McCain said to one agitated questioner, telling him to sit down. 'You've had enough time, sir. You've had enough time.'
McCain urged compassion. 'We are a
Judeo-Christian nation,' he said. He added that the border near Yuma is
largely secure, but he said smugglers are using the border near Tucson
to pump drugs into Phoenix.
McCain's second town hall meeting took place in Green Valley, south of Tucson.
A bipartisan group of senators —
including Arizona Republicans McCain and Jeff Flake — want assurances on
border security as Congress weighs what could be the biggest changes to
immigration law in nearly 30 years. Arizona is the only state with both
of its senators working on immigration reform in Congress, a sign of
the state's widely debated border security issues.
activists and elected officials say it's only natural for Arizona to
continue to take the forefront in the national conversation on
immigration after years of internal debate on the topic.
'There are 11 million people living here illegally,' McCain said. 'We are not going to get enough buses to deport them'
'No state in this country has had more experience with enforcement-only immigration laws than Arizona,' said Todd Landfried, executive director of Arizona Employers for Immigration Reform, which opposes the state's tough immigration laws.
While McCain defended his proposals to town hall meetings, Homeland Security Secretary Janet Napolitano toured the border near Nogales with the highest-ranking official at U.S. Customs and Border Protection, the incoming chairman of the Senate's homeland security committee and an Arizona congressman. Napolitano, Arizona's former governor, said afterward that comprehensive immigration reform will strengthen the nation's border against criminals and other threats.
Arizona gained international recognition as an epicenter of the U.S. immigration debate when it passed its tough anti-immigrant law in 2010. A handful of other states — including Georgia, Indiana, South Carolina and Utah — have since adopted variations of Arizona's law.
Arizona has the nation's eighth-highest population of illegal immigrants, according to the Pew Research Hispanic Center. In 2010, illegal immigrants represented roughly 6 percent of the state's population.
Arizona gained international recognition as an epicenter of the U.S. immigration debate when it passed its tough anti-immigrant law in 2010
Activists said Arizona's
anti-immigrant laws inspired many illegal immigrants to demand more
rights. Last week, some college students rallied outside Arizona Gov.
Jan Brewer's office for driver's licenses for illegal immigrants.
'They no longer are afraid to come and say, 'I am not able to vote, but I can make my voice heard, and they have to listen to me,'' said community organizer Abril Gallardo.
A report released in January showed the U.S. Border Patrol's Tucson sector remains the busiest along the U.S.-Mexico border. The Tucson sector accounted for 38 percent of all drug seizures and 37 percent of all apprehensions along the border.
Brewer said last week the border cannot be declared safe until the people living near it feel secure from drug and human trafficking.
But Democratic Congresswoman Kyrsten Sinema of Arizona told Latino and black community leaders at a Phoenix luncheon Tuesday that Arizonans need to spread the word on how much more secure the border has become.
'There are lots of folks who don't live in Arizona who have no idea what the border is like,' Sinema said.
VIDEO Sen. McCain argues with angry constituent over immigration
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Ask Health Experts and Get Answers to Your Health Question ASAP
yes have been tested and the tests came back positive for type 2..
ok thanks that explains why it has taken so long for me to get symptoms.. Is it possible that he is takening antiviral medicines as to why it has taken so long for me to get infected as well??
is it possible that i had the infection for some time but did not know and just now had a mild outbreak... If so what are the chances of my partner beening infected now??
I have been told it is more common that a man infects a women is this true..
no i was not told and i did not ask as i dont know what they are.. what are they..
how easy is it to infect other partners with unprotected sex??
i was certin that he infected me as it was only 2 weeks ago i got a few lumps it was so very mild and i did not get any other symtpoms at all... but now im wondering if i had the virus all the time.....
i have been reading all sorts of blogs on the net some say its easy to get infected from unprocted sex and some say it take a few times before you get infected sooo what is true how easy is it to get infected...
awesome thanks soooo much for your help... i now just have to wait for the IgM/IgG to get the turth now...Very muchly appreciated...


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Spell Keeper Silk Bag
by Fantasy Artist, Lisa Parker
Lisa Parker's Spell Keeper features the owl, in front of Celtic designed window among books, perched on a crystal ball as he holds a silver pentagram on chain. Striking art in which you will delight in storing your tarot or gems in.
100% Silk: Art Print is on one side; black silk on other side with drawstring closure.
Approximate Dimensions: 5 1/2 inches (14cm) x 7 3/4 inches (20cm).


text:
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LIGHTS, LANTERNS, CANDLES, Lamps, LIGHTED Props Decorations (195)
- Battery-Operated and Corded Electric Halloween Gothic Theme LIGHTS, Pirate Lanterns, Holiday Candles, LED Tealights, Haunted House Lamps, Special Effects Lighted Props, Light up Centerpieces, Spooky Chandeliers, Glow Light Sticks Light-Up Decorations
|Speaking Life Size ANIMATED RISING PHANTOM Reaper Horror Prop
|Spooky TALKING GIANT CORN STALKER Animated Halloween Haunt Prop
Creepy Life Size ANIMATED RISING PHANTOM Demon Grim Reaper Deluxe Animatronic Zombie Halloween Horror Prop Gothic Haunted House Speaking Greeter. New 6.5-Feet Tall Full Size Spooky Standing Display. Scary Light-up Sound Motion-Activated Talking Angel of Death Haunt Decoration
Creepy Animated SPOOKY TALKING GIANT CORN STALKER. This terrifying LIFE SIZE Deluxe Animated Scarecrow Jack-O-Lantern wire frame figure stands at an imposing 7-feet-6-inch tall and features a flickering-light-up Jack O' Lantern turning head and body.
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POSSESSED ANIMATED PROP Scary Gothic Horror Halloween Decoration-She's possessed! Designed to lay on her back on floor or table while shaking violently, moaning and screaming. Soundtrack CD included. CD player not included. All electric, life size prop Clothing may vary
Deluxe Latex LULLABY ANIMATED PROP Creepy Gothic Horror Halloween Decoration - Ghostly mom and child rocks back and forth singing softly a lullaby. Unnerving! Soundtrack CD is included. You provide chair and CD player. All electric! Clothing may vary.


text:
Amy was sitting outside in the rain, just outside of Tsuko's home, while all around her the city was smoldering from Tsuko's raging fire. Her encounter with Lady Thorn…or, as it turned out, the real Amy, had changed her outlook on life. Tails lied to her…he said that she had merely been sleeping for all those years, when in reality, she was just a clone…
Shadow took notice of the distressed girl, and took it upon himself to talk with her.
"Amy? You okay?" he asked as he sat down next to her.
"I'm not Amy…I never was, I never will be. You know it, too!" she replied.
"I don't give a damn. For all I care, you are the original."
"Well I'm not. Get used to the notion."
"I'll try."
Shadow was silent for a minute. Amy continued to sit there, the rain dripping down her head, face and every other part of her. He heard Tsuko emit a piercing, echoing scream.
"Listen to that, Amy…Tsuko is fighting for someone he has lost…someone very dear to him. And from the sounds of things, he's losing."
"So?" Amy said unemotionally.
Shadow was aghast. "So? So! SO? Listen to yourself! You've lost all will to go on, just because you find out you're artificially made! I was too!"
"Well, you aren't a clone. You're your own individual…I'm just a copy. Those memories of being saved…they were all lies!"
Shadow sighed and threw his hands up in frustration. "If this is how you're going to be, fine! I'm going to go help Tsuko. You obviously are too weak to be of any help."
Shadow started off back towards the castle without another word.
-- -- -- -- -- --
All the while, Amy sat there, pondering. As she tried to dig through her implanted memories, something came up that seemed real to her.
Amy stumbled down the street, crying her heart out, and holding her hand over her left eye. Turning into an alley, she made her way into a seemingly random apartment building, right past the landlord's office. She made her way down the hall, and knocked on Apartment 1-R. The door opened, and there stood Shadow, holding a dripping chili dog in one hand.
"Amy!" Shadow gasped. He dropped the chili dog, staining his shoe. "What happened?"
"Bad fall when I was with Sonic…he…just left me…"
"That must have been some fall…are you sure this isn't Sonic's doing? He's been awfully mean and angry lately."
Amy feebly revealed her eye. It was bruised severely.
Shadow winced a little. "Geez, I wish I could take you to a hospital, but I can't be seen in public. I'll do what I can, though."
Amy dried her tears and smiled. "Thanks…you're such a friend…"
"You did help me realize the good in me…are you sure Sonic isn't beating you?"
"I'm sure." Amy was silent for a moment. "Shadow?"
"I need a hug."
"Well…alright." He did so, and then briefly left the room to get his first aid kit. He never needed it, but these days, Amy sure seemed to need it.
"Shadow…if something ever happened to me, would you…save me?" Amy asked as she held a cold pack over her black eye.
"I…guess so…at least I'd try."
Amy's reminiscing was cut short by another scream of pain. But it wasn't Tsuko's this time…it was Shadow's. She then heard Sonic's maniacal cackling. Somehow, it stirred something inside of her…then it hit her.
Shadow has always been there for me. The promise he made fifteen years ago…that he would always be there if Sonic ever treated me badly…
Amy stood up, but did nothing. Another shriek of pain from Shadow pierced the air. The feeling within became stronger. But still she did nothing.
A third scream, this timechanged by something of a gurgle, issued forth. Amy's fear and doubt turned to anger. Her tears began to stream down her face, and energy began to course through her. The burning feeling in her forehead that happened a week before started again. Finally, the energy erupted fully, and one final thought crossed her mind…
Shadow protected me, Amy Rose, he made everything better. Even if I am not real…I can still help Shadow…
She faced the castle, and after staring at the sky for a minute, took off.
It's time to return those favors.


text:
Listen to free music by Tv Trax
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text:
Life is an eternal conquest of ideals and dreams unfulfilled.
Do not just gain wisdom, one must
use it .(Cícero)
It is better to conquer yourself than to win a thousand battles. (Buddha)
Fight for love is good, but to achieve it without fighting is best. (William Shakespeare)
If love is fantasy, ultimately I find myself in full carnival.
(Toquinho and Mutinho)
Win is not sufficient. We must learn to seduce.(Voltaire)
You know what is the reason for my smile everyday? The first word of that phrase.
In man, the desire begets love. In women, love begets desire. (Jonathan Swift)
In vengeance and in love woman is more barbarous than man.
(Friedrich Nietzsche)
True love has no happy ending, because true love never ends.
(Miley Cyrus)
Does not love the same woman twice.
(Machado de Assis)
Created by Antonio de Moura Rodrigues over 3 years ago


text:
When Apple's iPhone 6 Plus fell victim to the bendgate saga that had put several other similarly-sized gadgets on the limelight, we mulled over whether Apple would change anything in the upcoming model that would sort things, and here we are.
Future Supplier, a firm that sells parts of mobile phones and similar gadgets, just got hold of an iPhone 6S Plus shell that they think is sturdier than that of the original iPhone 6 Plus.
So if you feel you wouldn't spend a huge sum to buy a phone that bends, we can safely assume that Apple has heard you.
While a layman would hardly find any difference between the new case and the old one, seasoned Apple-watchers at Future Supplier noticed a change in position of the screw hole for the loudspeaker. They also said that the case looked and felt a lot stronger than the original iPhone 6 Plus case.
Apple is set to unveil its next range of iPhones in September as per its annual release cycle, unless the launch is either preponed or delayed by the Cupertino giant. Among the most plausible rumours are that the next iPhones would retain the physical characteristics of the existing ones but will feature several new changes in terms of internal specifications and functionalities.


text:
Willow Fork Drainage District's 2nd Birthday Bash
Central Green Birthday Bash!
May 9, 2015
Are you ready for fun? We have an amazing evening planned for you! Dancing Under the Stars with Cooper Wade and Pointe of Grace Dance Studio
- FREE Country Dance Lessons 6:00 pm
- Country Artist, Cooper Wade, 6:30 pm
- Curt Miller Magic and Comedy 6pm (Walk Around Magic)
- Giant Slide and Bounce House 6pm-8pm
- Face Painting
- Vendor Booths
Event Files
Sat, May 9, 2015
6 p.m. - 10 p.m.
(GMT-0500) US/Central
Central Green Park
23501 Cinco Ranch Blvd
Katy, Texas 77494
United States


text: On Wed, 2006-03-08 at 23:26 +0530, Rahul Sundaram wrote: > Jeremy Katz wrote: > >On Wed, 2006-03-08 at 08:29 -0500, J. Hartline wrote: > >>Provide fedora and root usernames with blank passwords. Security is not > >>a concern for a Live CD > > > >No! Security is very much a concern here. We need to make sure we > >don't start a spread of worms from our live CD. > > > Auto login to a non administrative user called Fedora. How is that bad > for security? If you have ways to log in remotely, then it could allow for people to log in and start running, eg, eggbots or any of a number of other things if there's a blank password. So it's not necessarily that auto-login to a non-administrative account is bad (it probably does make sense). But just that thinking "oh, security isn't a concern for a live cd" is not wise. Jeremy


text:
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text:
Mascapone Chestnut Yogurt Mousse
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READY IN: 10mins
Recipe by InnerHarmonyNutriti
This recipe requires only a few minutes of mixing. Chestnut spread is often used in France and can be purchased on Amazon. The spread is quite sweet so you don't need to add extra sugar unless you have a real sweet tooth. If you don't wish to use rum, I suggest adding vanilla extract instead. For more healthy gluten-free, pesco-vegetarian recipes, please visit my blog, www.InnerHarmonyNutrition.com.
See Discussion:
Ingredients Nutrition
- 8 ounces mascarpone cheese, at room temperature
- 2 3⁄4 ounces chestnut puree (spread)
- 150 ml plain yogurt
- 1 tablespoon rum
- 1 teaspoon cocoa powder, for dusting
- 1 teaspoon chocolate shavings (optional)
Directions See How It's Made
- In a bowl, mix the first four ingredients using a hand mixer.
- Divide the mouse into 4 or 5 dishes.
- Dust some cocoa powder and chocolate shavings on top.
- Refrigerate. Infuse love and serve!