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Houston Felony Lawyer James Sullivan defends clients accused of all felony crimes. Serving Greater Houston since 1994. Call (281)546-6428 for a free phone consultation.
Houston Juvenile Lawyer Defends Victims of Bullying that Fight Back
Harris County Juvenile Attorney James Sullivan strongly believes that child and young adult victims that stand up to a bully, especially in school, should be applauded, not arrested.
Bullies generally target children they view as being weaker, smaller or socially awkward. Children with mental health disorders, such as autism spectrum disorder, are especially vulnerable to being bullied.
Effects of Being Bullied
According to Psychiatry, a peer-reviewed journal of the National Institute of Health,
In the aftermath of being bullied, victims may develop a variety of psychological as well as somatic symptoms, some of which may persist into adulthood. Psychological symptoms may include social difficulties, internalizing symptoms, anxiety, depression, suicidal ideation, and eating disorders (i.e., anorexia or bulimia nervosa). Somatic symptoms may include poor appetite, headaches, sleep disturbances, abdominal pain, and fatigue.
For many victims, the bullying lasts for weeks, months and even years before the victim finally has enough and musters up the courage to fight back. And then, instead of being commended for standing up to the bully, the victim is often victimized again by the school district police department which labels him the aggressor and arrests and charges him with Assault causing bodily injury.
The parents and grandparents of today's youth grew up in simpler times when fighting in school was handled by the school, usually by paddling. School principals understood that when victims finally stood up to bullies and fought back it is was huge for their self-esteem and the bullies stopped picking on them.
Zero Tolerance Policies Should be Reformed
Texas' "zero tolerance" policies should be changed so that the victims of bullying who fight back or lash out are not arrested and charged in court. Even psychology experts agree that reforms need to be made. According to the American Psychological Association Zero Tolerance Task Force,
Zero tolerance policies as applied appear to run counter to our best knowledge of child development. By changing the relationship between education and juvenile justice, zero tolerance may shift the locus of discipline from relatively inexpensive actions in the school setting to the highly costly processes of arrest and incarceration. In so doing, zero tolerance policies have created unintended consequences for students, families, and communities.
Effect of Current Laws
Unfortunately, until these zero tolerance policies are changed, victims of bullying continue to get into serious trouble for fighting back or lashing out. In addition to in or out of school suspension or placement at an alternative school, these victims frequently get charged with Assault and wind up in juvenile district court.
A juvenile delinquency charge puts the student at risk of a permanent juvenile record that could negatively impact his future, such as acceptance into top universities or enlistment in the military. Attorney James Sullivan fights for these students so that their future is not ruined by a juvenile or criminal record. Sullivan defends children and young adults facing Assault charges from all of the school districts in Harris County.
If your son or daughter is facing Assault charges in the 313th, 314th or 315th District Courts of Harris County, it is important to hire an experienced juvenile defense attorney.
James G. Sullivan and Associates
Attorney James Sullivan has been defending children and adults accused of Assault in juvenile and criminal courts since 1994. Sullivan is also an expert in juvenile law, having been Certified in Juvenile law by the Texas Board of Legal Specialization since 2004.
To discuss your child's case, contact James Sullivan at 281-546-6428 for a free phone consultation.
Posted by Jim Sullivan
12345 Jones Road, Suite 288, Houston, Texas 77070
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"A new commandment I give to you, that you love one another. Just as I have loved you, you also must love one another. By this everyone will know that you are my disciples, if you have love for one another." John 13:34-35 NHEB.
Previous Bible study lesson: 613 Commandments #5.13.
1) What is the New Commandment?
2) Why does the Law of Christ exist?
3) How do you obey Jesus' Commandment?
There is so much confusion in Christian life about the use of the Ten Commandments.
Why is there one set of commandments in the Old Testament but suddenly there is this thing called "Jesus' Commandment" in the New Testament?
This will be an incredibly important Bible study for your Christian walk. You'll gain knowledge that's hard to come by and hidden from most people, theologians included.
This study isn't for the spiritually meek or cowardly. Pull up your boot straps, tighten your belt of truth, and put on your helmet of salvation. You're entering the spiritual battle zone against a ticked-off satanic army.
I hope your spiritual life will be transformed like mine has from this study. Let's get started.
The Old Covenant, aka Old Testament and the Law, was glorious and used greatly by God in growing his people's knowledge of their coming Messiah. The Law is good, but, sin stops us from obeying it. We are fallen and the Law tells us so (Romans 7:14-16).
The Old Covenant ended in death and condemnation, though, because the sinful nature of man prevented everyone from obeying its laws. God himself saves, the Law does NOT.
The Old Covenant wasn't intended by God to save people's souls. It only brought knowledge, conviction, and guilt, which finally resulted in a curse (Romans 7:11-13). That is one of the Law's purposes - to reveal God's curse on sinful humanity (Galatians 3:10-11).
Most assuredly the Law was important, and still can be, with its goal being Jesus Christ (Matthew 11:13). The Old Covenant's purpose was to usher in the Messiah. The Covenant directly pointed to Jesus Christ (John 5:39-40; Romans 3:25-26). True spiritual Jews looked to the coming Messiah to save them, and when he came he did, in fact, save them from condemnation and eternal death (John 3:17; Hebrews 9:15). Hold this thought inside your soul while we focus more on the Law.
For a moment, let's look at what the Old Covenant Law did not do. The Old Covenant didn't change a sinner's heart. It didn't forgive and finally remove one's guilt. It didn't permanently redeem, justify, or pardon anyone. The Law does not save souls, but it is a critical part of God's plan of redemption (Romans 5:20). The New Covenant accomplishes all these things the Old Covenant wasn't meant to do.
What the Old Covenant could not accomplish the Son of God did by fulfilling it himself, because he alone could obey the law perfectly. He succeeded when all others failed. Jesus Christ came to earth to fulfill the Law (Matthew 5:17-20), which he did. Jesus Christ ALONE appeased the wrath of God when ALL humans were unable and unqualified.
We are ALL still unable to perfectly obey God's law, and that includes both you and me (Romans 3:20). On top of all that, God's full Law included the need for a sacrificial death for humanity's sins, of which Jesus also fulfilled, thereby appeasing the Father (Hebrews 10:12-13).
The Apostle Paul tells us the Old Covenant, aka Old Testament, has been replaced with the New Covenant, aka New Testament. The author of Hebrews says the same, "When God speaks of a "new" covenant, it means he has made the first one obsolete. It is now out of date and will soon disappear" (Hebrews 8:13; 10:1; 10:9).
But, does the new law sound much different than the old law, specifically the Ten Commandments, which mostly deal with morality?
The Law of Christ (1 Corinthians 9:21) calls us to a faith in action that loves others, forgives others, and shares the burden of others. Listen to this command, "Bear one another's burdens, and so you will fulfill the Law of Christ" (Galatians 6:1-3).
Jesus' Commandments call us to love others so much that we help them through their sin, and back into beautiful fellowship with God and others. We're not to stop at our neighbor's sin, though, meaning we're to share their burdens of pain, distress, persecution, sickness, loss, and grief. We are to help one another through all of life's many difficulties.
The Law has ALWAYS been to love God and love one's neighbors. Why are we so surprised about all this? The Law's purpose has NEVER changed! (read Deuteronomy 6:5; Leviticus 19:18; 2 John 5). The Law points us to Christ (Romans 3:19-20), who in turn, points us to love God and other people.
What HAS changed with the New Covenant is that we now have the Spirit of Christ living in the hearts of true believers (Ezekiel 36:26-27). The Spirit empowers us to live more intimately with God and at a higher level of obedience to him. We're now greatly motivated towards obedience by the love shown in Jesus' life and death. He died for us, so now we live for him (Titus 3:4-8). This all makes perfect sense, doesn't it?
So, what use is the Old Covenant now? What is edifying about reading the Old Testament? First, We learn about God. We learn about humanity. We also learn from the Hebrews' relationship with God (1 Corinthians 10:11; Ephesians 2:7). Secondly, we stand in amazement of the Old Testament prophecies and their fulfillment in Christ Jesus. The Old Covenant, therefore, builds our foundational faith in Jesus Christ (Luke 24:25-27). Finally, the Law exists for the rebellious person, to give him or her knowledge and conviction of sin (1 Timothy 1:9). The Law, in fact, still exists for pointing a sinner to Jesus Christ (Galatians 3:19; Romans 3:20; 7:7-8).
Can YOU see Jesus? Can YOU now see why our focus has shifted to Jesus' Commandments?
Let's turn our attention to learning more about the Law of Christ, his New Commandment (John 13:34-35).
If the Old Covenant is explained throughout the Old Testament, then it goes without saying that the New Covenant is found throughout the New Testament. God used the prophets to write his story and his words in the Old Covenant, and Jesus used his apostles to write his story and his words in the New Covenant.
The Old Covenant pointed the Hebrews' hope towards the New Covenant and its Messiah (Matthew 11:13; Romans 3:25-26). Don't you love this knowledge? I told you this would be a terrific study!! It gets even better, believe me.
Pure belief in the two Testaments of the Bible is tantamount to belief in God. To deny the Bible as straight from God's throne is: defiant unbelief (Matthew 22:29), proud heresy (Matthew 15:6), and an eternally damnable activity (Mark 8:38). Humble repentance is in order if you find yourself in such a terrible place of unbelief.
Jesus' commandments are found throughout the entire New Testament. Christ fulfilled the Old Covenant, and then ascended to heaven to be our High Priest (Hebrews 10:12-13). Jesus Christ ascended but left his Church the Holy Spirit and his word, which is found in the Bible (John 16:5-10).
This New Testament explains the mystery of the New Covenant between God and humanity (1 Corinthians 2:7). It's the written documentation of the fulfillment of the beautiful redemption story. Nothing can be added to the Scriptures, and nothing can be taken away (Revelation 22:18-19). God forbid that you allow the biases of modern science, or some cult's extra-biblical teachings, to shipwreck your pure belief in God's Bible. That ultimate failure will lead to your soul's destruction.
We have Christ's words now firmly established throughout the New Testament. Christ's Law (aka the Law of Christ, Law of Liberty, Royal Law, or the New Commandment) is what Christians are expected to live by (2 Timothy 3:16-17).
To truly follow Jesus Christ we MUST follow the New Testament. We have a severe warning given us to protect us from damnable hypocrisy (Hebrews 10:26-31). Don't dare say you follow Christ when you trample Him and his word under your feet. Flee from your dark sin towards the One who brings the Light (John 12:35-36).
Before going further, please understand that faith alone, in Jesus Christ alone, saves your soul (Ephesians 2:8-9). Grace alone, from Christ alone, keeps you saved (Galatians 3:1-3).
Obeying Jesus' commandment, and believing his entire word, displays your heartfelt thanks to God for your salvation. You don't gain or keep your personal salvation by your obedience or goodness, period. Good works and persistent, properly motivated obedience PROVES your faith to God and the watching world. Lack of good works, love for others, and disobedience to the Bible PROVES you have deluded yourself through misguided faith.
Salvation is by faith, through God's grace, not of works. You cannot boast in your goodness, efforts, words, or works (Galatians 3:21-22). Jesus wants all of your heart, so give it to him. Your heart and actions will reveal to you if he has set you free from sin (John 8:34-37).
Some people say that there are 39 of Jesus' commandments found in the New Testament. I find the number of his commandments very subjective, and don't feel the need to "go there," i.e. to specifically number Jesus' commandments. Here are some reasons why.
First, "listing" Jesus' commandments may lead to the practice of legalism (2 Corinthians 3:6). The list-follower may fall into the devil's trap of strictly following the commands and concluding that God loves him or her because of obedience (Romans 7:6).
Secondly, Jesus' commandments aren't just from his own mouth, but also written in the epistles of the New Testament. Jesus used the apostles to write his words, so the New Testament is full of his commands. Finally, Jesus' commandments are spiritual, and mean different things to different people at different times. They are not static and concrete, but meet people at their spiritual level (2 Cor 3:3).
However, with these qualifications on the table, I'd like to offer a "partial" list of Jesus' commandments, starting with the two summary commandments. (See bottom of this page for a link to a list of 1,050 New Testament commands).
The First and Greatest Commandment: "You must love the Lord your God with all your heart, all your soul, and all your mind" (Matthew 22:37-38).
The Second Greatest Commandment, aka The Golden Rule: "Love your neighbor as yourself" (Matthew 22:39; 7:12).
These Two Great Commandments are the basis of ALL of God's Laws (Old Covenant) and ALL of the Laws of Christ (New Covenant). All biblical law, both Old and New Covenants fall under these two mighty, over-arching Laws of God and Christ. (Deuteronomy 6:5; Leviticus 19:18; Matthew 22:40; 2 John 5).
"Repent of your sins and believe the Good News!" (Mark 1:15).
"Believe in the One He has sent." (John 6:29).
"The Scriptures cannot be altered." (John 10:35).
"You must be born again." (John 3:7).
"If any of you wants to be my follower, you must turn from your selfish ways, take up your cross daily, and follow me." (Luke 9:23).
"Stop sinning!" (John 5:14; 8:11).
"Stop complaining about what I said." (John 6:43).
"I am the Way, the Truth, and the Life, no one comes to the Father but through Me." (John 14:6).
"Anyone who obeys my teaching will never die!" (John 8:51).
"Love your enemies! Pray for those who persecute you!" (Matthew 5:44).
"Forgive those who sin against you, so your heavenly Father will forgive you." (Matthew 6:14).
"Don't store up treasures here on earth... store your treasures in heaven." (Matthew 6:19-21).
"You cannot serve both God and money." (Matthew 6:24).
"Do not judge others and you will not be judged. For you will be treated as you treat others." (Matthew 7:1-2).
"Whoever divorces his wife and marries someone else commits adultery against her. And, if a woman divorces her husband and marries someone else, she commits adultery" (Mark 10:11-12).
"You must not commit adultery." (Mark 10:19).
"I am sending you out as sheep among wolves." (Matthew 10:16).
"You live by the sword you shall die by the sword." (Matthew 26:52).
"Go and make disciples of all the nations, baptizing them in the name of the Father, and the Son, and the Holy Spirit. Teach these new disciples to obey all the commands I have given you." (Matthew 28:18-20).
"Everyone who endures to the end will be saved." (Matthew 10:22).
This list of Jesus' commandments is but a small taste of what he commands in the New Covenant. Being Christ's disciple isn't something to be taken lightly. Count the costs, my friend. Think eternally and give up your personal rights to God. Become a slave of Jesus (Romans 6:20-22) and enjoy real freedom.
Jesus expects you to love these commands, doing your best to follow them. Remember the main point, Jesus fulfilled the commandments of God perfectly, therefore you must trust him for eternal life. That is your main and first assignment as his disciple - Trust in Jesus (John 6:29).
Trusting God through Christ offers you forgiveness for all your sins and it grants you eternal life (John 5:24). Loving his commands and doing your best to obey them reveals a humble, childlike faith, which is the attitude Jesus expects from his disciples (Matthew 19:14).
How do the 10 Commandments relate to the New Covenant? Are they still respected by Christ? These are good questions and the answers vary greatly amongst Bible scholars.
Jesus and his apostles reaffirmed the Ten Commandments in the New Covenant. Really, they amplified them. The new covenant is called the "perfect law of liberty" which contrasts with the old covenant of death (Galatians 2:19), i.e. the Ten Commandments and God's entire Law.
View the Ten Commandments as physical commands, because God expected physical obedience (Deuteronomy 6:25). Then, view Jesus commandment as spiritual, because the Holy Spirit enables a heart change before God expects physical change. The new covenant brings the believer a new heart before the believer learns new ways of living (Romans 8:2-3).
Following the Law of Christ releases us from judgment and eternal death (James 1:25; 2:8-12), which the Ten Commandments never intended to do.
1st Commandment: "You shall have no other gods before Me." Exodus 20:3, restated in Matthew 22:37-38 and Acts 14:15.
2nd Commandment: "You shall make no graven image." Exodus 20:4, restated in 1 John 5:21.
3rd Commandment: "You shall not take the name of the Lord your God in vain." Exodus 20:7, restated in James 5:12.
4th Commandment: "Remember the Sabbath Day, to keep it holy." Exodus 20:8-11, restated spiritually in Matthew 11:28-30. Sabbath means "rest" and the New Covenant calls us to rest in Christ from our efforts to gain salvation. Jesus' rest is eternal (Hebrews 4:1-11).
5th Commandment: "Honor your father and your mother." Exodus 20:12, restated in Ephesians 6:1.
6th Commandment: "You shall not murder." Exodus 20:13, restated in 1 John 3:15.
7th Commandment: "You shall not commit adultery." Exodus 20:14, restated in 1 Corinthians 6:9-10.
8th Commandment: "You shall not steal." Exodus 20:15, restated in Ephesians 4:28.
9th Commandment: "You shall not bear false witness against your neighbor." Exodus 20:16, restated in Colossians 3:9-10.
10th Commandment: "You shall not covet." Exodus 20:17, restated in Ephesians 5:3.
Most of these laws are moral laws, first given in the Old Covenant. The indwelling Spirit of Christ (New Covenant) brings Christians to a higher level of obedience to God's Law, now known as the Law of Christ. The Ten Commandments have essentially been "filled out" by Jesus' Commandments.
Jesus wrote between the lines of the 10 Commandments! (John 8:4-8). They are now complete, and Jesus' Spirit gives us the knowledge, motivation, and power to follow them! The New Testament contains Jesus' New Commandment.
Jesus stated, "If you love me you will obey my commandments" (John 14:15). Loving Christ is loving his commandments. Jesus' commandments are light, and they are easy to follow (Matthew 23:3-4), they are gratifying to the soul of a person who longs for love, truth, reality, righteousness, and eternal life (John 6:35).
Loving Christ equates to loving God and loving other people. Loving Christ gives up your so-called personal rights, and serves God and other people. Loving Christ is total commitment to Jesus.
Trying to obey these commandments on our own is futile and fruitless (Galatians 3:10-11). Don't even try (Romans 3:21). The place to start obeying Christ is on your knees, face buried in your hands, begging God for mercy. Ask for the Holy Spirit, the power of God, so that you can believe in Christ and follow the Scriptures (Romans 3:22).
Upon Christ's response to your plea, you'll notice a warm, heavenly peace wash through your body (John 14:27). The living water from heaven then begins removing your old worldly loves (John 4:14). You'll soon notice a deep love for Jesus and other people. The experience is unmistakable.
Jesus would never tell us to do something that he wants us to do in our own power. Faith is required for all true obedience, and God provides all the faith that we possess (Romans 10:17). Following the law of Christ is a call to faith and action.
Faith begins this journey in the Spirit (Galatians 3:5); and by the Spirit of Christ believers produce the fruit of love (Galatians 5:22-23); and finally, through love we fulfill the law of Christ (Galatians 6:2).
It is really that easy. Trusting Christ produces fruit, which results in love, thus fulfilling the Law of Christ (John 5:24).
You won't be perfect, in fact, you'll completely fail at times. You'll really blow your testimony, too. However, because of the finished work of Christ and his faithfulness to his true disciples, you can be forgiven and renewed. Repentance keeps God's mercy upon you, renewing your faith in God. God is so full of amazing grace.
Trust in Christ now, and eternally rest from your hopeless efforts to please God through good deeds.
Next Bible study lesson: Catholic Ten Commandments #5.15.
1) Can a true Christian dislike certain people and please God in the process? Discuss.
2) Are Christians misguided who prefer studying the Old Testament more than the New Testament? What might, but not necessarily, this preference indicate about their spiritual maturity?
3) Do pew-sitters need to fear Jesus' Commandments? Why or why not?
There is therefore now no condemnation to those who are in Christ Jesus. For the law of the Spirit of life in Christ Jesus made you free from the law of sin and of death. For what the law could not do, in that it was weak through the flesh, God did, sending his own Son in the likeness of sinful flesh and for sin, he condemned sin in the flesh; that the ordinance of the law might be fulfilled in us, who walk not after the flesh, but after the Spirit. Romans 8:1-4 NHEB.
But, beloved, we are persuaded of better things for you, and things that accompany salvation, even though we speak like this. For God is not unrighteous, so as to forget your work and the love which you showed toward his name, in that you served the saints, and still do serve them. We desire that each one of you may show the same diligence to the fullness of hope even to the end, that you won't be sluggish, but imitators of those who through faith and patience inherited the promises. Hebrews 6:9-12 NHEB.
Dear Lord, I get it now. I understand the nature of your Law. You're holy, and we're not, and have no hope of becoming holy by your Law. The Law shows us that we're hopelessly sinful and unable to please you. But, thank YOU, Lord, for sending Jesus to come and fulfill that Law. Thank YOU for applying Jesus' life to our account. Thank You that the Law of Christ calls us to a higher place. Jesus' laws are easy, edifying, peaceful, and satisfying. The Holy Spirit gives us a love for your Law, and he gives us the power to obey. Lord, forgive my failures, my sin, my shame. Help me to love ALL my neighbors, and carry their burdens. Through the blood of Jesus, standing on your promises, Lord, I know you will enable me. You are truly a great, great God. In Christ's Name, Amen.
Go in peace, my friend. I hope you enjoyed the Bible study lesson.
Share on Facebook below and click for your Free Printable Bible Study Lessons: Jesus' Commandment #5.14. Click for your free printable List of the Ten Commandments. Please Donate to GG.com!
New! List of the 1050 New Testament Commandments.
Special Note: I'm grateful to Big Book Media & Free Bible Images, stock.xchng, Morgue Files, and Stock Photos For Free for the use of their photos for Jesus' Commandment Bible Study Lesson.
Don't miss this incredible video showing you how to apply Jesus' Commandments to evangelism!


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K2 Announces Partnership with UiPath to Expand Robotic Process Automation Capabilities
1/1/2019 3:00 PM (UTC)
Bellevue, WA, Oct. 03, 2018 (GLOBE NEWSWIRE) -- K2, the leader in digital process automation, announced today a partnership with UiPath, a leader in robotic process automation (RPA), to integrate their two platforms and enable full lifecycle business automation.
The new native integration between K2 and UiPath provides the flexibility you need to use robots to start or update inflight K2 processes or have K2 call a UiPath robot to fully automate tasks. This full lifecycle approach allows humans and RPA robots to work together seamlessly, ultimately reducing costs and improving accuracy, compliance and agility.
"With the meteoric rise of process automation in the market, our customers are looking for ways to automate repetitive tasks so that their employees can focus on more critical aspects of business," said K2 CEO, Evan Ellis. "We're thrilled to partner with a leader in the space to bring our capabilities together and jointly expand business automation."
The emergence of low-code digital process automation and RPA platforms have enabled a fast-growing number of companies to digitally transform. Forrester found that accelerating digital business transformation now outweighs cost reduction as a primary focus of process improvement, in a survey of over 300 business and technology decision makers.iHowever, manual processes continue to plague customer experiences.
"Manual processes cripple customer journeys by thwarting a seamless experience. Part of any overall automation effort involves your IT and business teams assiduously identifying and eliminating manual processes. You can automate much of that work with more straightforward deterministic tools like emerging low-code development products that make previously complex development and integration far simpler," wrote Forrester Research Vice Presidents and Principal Analysts, Rob Koplowitz and Sucharita Kodali, in a recent report. "You'll also want to explore robotic process automation (RPA) to automate tedious human-centric tasks like copying data from one system into another because back-end automation is too costly or complex."ii
"We're particularly excited about this partnership because K2 has built a native integration with our REST API and isn't relying simply on screen-scrape integration of a form. Customers will be able to bring humans into the loop effortlessly through K2, and when the human completes the task at hand, the process can be sent back to UiPath robots to keep things running," said UiPath VP of Product and Strategic Technology Alliances, David Marcus.
K2 customers today only need their UiPath credentials to integrate the two platforms for both inbound and outbound communication. To learn more and request a free trial, please visit K2.com.
About K2
K2 is the leading low-code, cloud-based digital process automation (DPA) platform for enterprises seeking to rapidly and intelligently create modern process applications, automate workflows and transform their business. With K2, thousands of organizations worldwide and 30 percent of the Fortune 500 have taken control of their business processes to increase visibility and improve operational efficiency. Discover what you can accomplish when you connect your people, processes and applications at K2.com.
About UiPATH
Built for both business and IT, UiPath is the leading platform for enterprise Robotic Process Automation (RPA). More than 1,800 business customers and government agencies use UiPath's Enterprise RPA platform to rapidly deploy software robots that perfectly emulate and execute repetitive processes, boosting business productivity, ensuring compliance and enhancing customer experience across back-office and front-office operations.
With a thriving RPA developer community of more than 250,000 worldwide, UiPath is on a mission to democratize RPA and support a digital business revolution. Based in New York City, UiPath's presence extends to 14 countries throughout North America, Europe and Asia. The company is backed by over $400M in funding from Accel, CapitalG, KPCB, Credo Ventures, Earlybird's Digital East Fund, Sequoia Capital and Seedcamp.
______________________________________________________________________________________________________________________________________________________________________________________
i The Growing Importance of Process to Digital Transformation. Forrester Research. May 8, 2018
ii AI in Retail? Let's Start with Automation in Retail. Forrester Research. August 9, 2018
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Using a dumpster will save people a tremendous amount of time. Garbage is usually thrown straight into the bin, rather than piled up and brought to the dump. There is no quick trip to the actual dump. By eliminating the need to generate there, folks save time. This time can be used to earn income, rather than stand.
For people who tend to be anticipating starting a major design or demolition venture, or even a venture that includes both of these operations, it is quite achievable, highly very likely in fact, that many debris are going to be generated.
Removing bathroom lighting fixtures and drywall can be a messy and tiresome project. A new do-it-yourself project can easily leave an enormous mess that won't have been regarded as. Regular rubbish pickup inside the community will demand the deteriorating of the drywall and positioning it straight into bags. It'll likewise require that the bathroom lighting fixtures be got rid of off in respect to the township's laws. This can take time and need that your old, stained shower sit away from your home even though it waits with regard to trash pick up. This is unneeded when you call for a Rent a Waste Management Dumpster in Peel. The size of the actual dumpster will be driven by the size of the actual project and the cost is driven by the amount of rubbish disposed of. A new Rent a Waste Management Dumpster in Peel is affordable and offers a quick implies to remove extra waste during a bathroom renovation.
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People want to keep their residence and surroundings clean and clean. Huge amount regarding trash is definitely generated in house renovation in addition to construction tasks. These waste materials cause trouble at your job location. To get rid of the clutter formed at your job location, you've to find the ideal Rent a Waste Management Dumpster in Peel service company.
A lot of people look at renting a dumpster as a possible expense. This specific cannot be rejected, as it is expensive to borrow the dumpster from a corporation. However, persons rarely think about the savings which using a dumpster provides. Businesses usually use them, mainly because dumpsters are the most efficient way to get rid of a large level of garbage. Yet, the dumpster can also ease the strain upon an individual's pocketbook. Here are three ways dumpsters save persons money when they accept large tasks.
Roll away dumpsters can typically be dropped off at in any site you desire providing it isn't creating injustice to someone else in the neighborhood. It is vital to know the correct sizing and time frame the dumpster will be needed so you can tell your neighbors as well as other businesses before hand. I have to claim, roll away dumpsters aren't one of the most eye catching device so allowing your fellow neighborhood friends know will be again essential. The trash corporation in which you choose will then generate a time to sometimes pick up as well as replace the actual dumpster should you deplete all of your space.
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Maybe you have bought the fresh new flooring and it is included and the defending plastic will require to be given away in the dumpster. When the drywall is carried out you can mount your completely new carpeting or maybe have the carpeting layers deliver the results for you.
Acquire associated with the Rent a Waste Management Dumpster in Peel, AR supplier, who updates the customer advance about the items which can be disposed into a dumpster, charges of the Rent a Waste Management Dumpster in Peel, AR and terms and conditions with the disposal. When you get associated with a corporation that can give quality providers at affordable cost, you may realize that trash removal upon completion of your projects was never that easy and that affordable.
Actions include: Evaluate which size dumpster you'll need. Do you need the whole sized or even a small dumpster. Look at the amount of trash that will need to have to be taken off by the completing the undertaking. Determine if you need a commercial or residential rental. An industrial rental may be needed should your contractor is progressing a huge redevelopment and is removing bricks and wood and so forth. Online online catalogs provide all of the answers you will require for selecting the right size and type. If you live in the Washington DC region you can need a Maryland Rent a Waste Management Dumpster in Peel, AR; you may also go online and an order or maybe call. A quick removal of the complete dumpster is available. Ones construction approach can be made much easier with a Rent a Waste Management Dumpster in Peel, AR.


Prepare for every presentation you make—and remember that every time you open your mouth or prepare a written communiqué, it's a presentation. Think about the two or three most important things you want people to take away, and keep your message focused on just those. Remember: short sounds confident.
Do your homework. My 7th grade science teacher, Hans Selye, used to tell us, "Chance favors the prepared mind." I never forgot that. (And that was quite a while back!) Regularly reading professional journals, newspapers, and magazines not only keeps your mind agile and open to new ideas, it also provides you with the ammunition you need to support your positions. Use the information you glean in making a pitch or a point.
Brainstorm ideas with people who think differently than you. If you know you're an "idea person" who has difficulty communicating the tangible benefits of your ideas, talk to someone who's good at it and ask for help in synthesizing your concepts for crisp communication. Conversely, if you tend to get bogged down in the weeds, look to those who are always coming up with new ways of approaching old problems for help in assessing the bigger picture.
If you like these tips, you'll love Urban Wealth Management's webinar on November 9th: "Want the Corner Office?" Dr. Lois Frankel will spend a half-hour with us giving us more tools and tips for getting what you most want at work and in life. You can also read more Dr. Frankel's interview with Diane Manuel on our blog. You can also hear Dr. Frankel in person at the Women's Career Conference of Pasadena, benefitting BloomAgain Foundation, on Saturday, October 28th.
Dr. Lois P. Frankel, President of Corporate Coaching International, is a bestselling author, executive coach, and an internationally recognized expert in the field of leadership development for women. She has appeared on Larry King Live, The Tavis Smiley Show, The Today Show, CNBC, and PBS to discuss her New York Times bestselling books, Nice Girls Don't Get the Corner Office, Nice Girls Don't Get Rich, and Nice Girls Just Don't Get It. Her other books include See Jane Lead and Stop Sabotaging Your Career. Lois's work with women and girls has been recognized with the Woman of the Year Award from the Los Angeles Commission on the Status of Women, Maybelline New York's Women Who Empower through Education award, and an honorary doctorate of Humane Letters from Phillips Graduate Institute.


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With partisans in the US arguing over whether Antifa or supporters of President Trump are to blame for the storming of the US Capitol, leaders of the Islamic State terror group are reportedly downcast that no one has even thought to accuse them of leading the attack. "Five years ago, if someone lit a firecracker in a Hooters parking lot,...
Recruiting Kanye Not Worth It, ISIS Decides
Despite the public relations boost it would give to an organization desperately trying to stay relevant, Islamic State officials have decided not to recruit rapper Kanye West to the terror group. "Honestly, we could probably get him to sign on as our spokesman in 15 minutes," ISIS leader Abu Ibrahim al-Hashimi al-Qurashi told The Mideast Beast, noting West's dramatic vacillation...
ISIS Doctor Loses License for Actually Saving Life
Calling him "disgraceful" and "an infidel," ISIS spokesman Abu Hassan al-Muhajir has announced the disqualification of Dr. Abu-Bakhr's license to practice medicine. The decision to revoke his license comes after the surgeon provided life-saving intervention to a little boy suffering from tuberculosis. "Abu-Bakhr intentionally defied the protocol set in place to prevent this type of incident," the spokesman said. "How...
Israel, Politics, War, World
UN Accuses Israel of Committing Genocide Against Coronavirus
With Israel leading the world in coronavirus vaccination rates, the United Nations General Assembly has passed a resolution condemning Israel for its ongoing genocide against COVID-19 virions. "The illegal apartheid state of 'Israel' is once again committing crimes against humanity, ethnically cleansing innocent coronavirus particles from their homes in the human body," the resolution states. "We call on the occupation...
God lists "Mideast Peace" as New Year's Resolution for 2021st Time
After another year of war between Middle East countries and factions, and a global pandemic, God has publicly tweeted that this year is going to be different. "2020 didn't go as planned. But new year, new me! In 2021 I vow that there will b no more mideast wars. No promises on Covid. ur fault 4 not wearing a mask....
Saudi Government and COVID Come to Agreement on Destruction of Yemen
In a ground-breaking first for medical science, the Kingdom of Saudi Arabia and COVID-19, the disease caused by SARS-CoV 2, have agreed to share responsibility for the devastation of Yemen. A spokesman for the Kingdom commented, "We've been at this for five years, and to be frank it's not exactly been 'Mission Accomplished'. It appears that a campaign of precision...
In an arms deal that could change the balance of power in the region, the United Arab Emirates has agreed to buy 100 million Twisted Tea cans from the US. The UAE, which was previously looking to purchase 50 advanced stealth F-35 jets, decided to instead invest its defense budget in the alcoholic beverage after seeing a video in which...
Sports, War
Trevor Lawrence Joins ISIS to Avoid Playing for Jacksonville
With the Jacksonville Jaguars clinching the top pick in next year's NFL draft, Clemson quarterback and top prospect Trevor Lawrence announced that he will move to Syria and join the Islamic State to avoid playing for the Jaguars. Lawrence, widely considered the best quarterback prospect in a generation, acknowledged that he was not looking forward to living in the Syrian...
Iran to Nuclear Inspectors: "Sorry, All Our Reactors have COVID-19!"
While the International Atomic Energy Agency (IAEA) is responsible for inspecting all of Iran's nuclear sites, the process has proven difficult, especially since the outbreak of COVID-19. What many nuclear inspectors have found surprising is the fact that every single nuclear site is reportedly testing positive for the virus, rendering them impossible to investigate. A chief IAEA nuclear inspector described...
Entertainment, War
Iran Releases 'A Christmas Story' Remake in Which Little Ali Asks Santa for a Nuke
Looking to tap into Americans' nostalgia for Christmas movies, the Islamic Republic of Iran has released a modern remake of the holiday classic 'A Christmas Story.' But instead of nine-year-old Ralphie Parker, the main character of the remake is 81-year-old Persian boy Ali Khamenei. And rather than asking Santa for a BB gun, Little Ali just wants a military nuclear...
Culture, War
Putin deploys S-300 Santa Claus Tracking System to Syria
According to the Russian Interfax news service, just in time for the holidays, Russia is planning to deploy an additional S-300 'Santa Tracking' surface to air missile system to Syria. The weapon will be capable of tracking Santa Claus across Syrian airspace. The Russian Ministry of Defense (MoD) insists that the system is nothing for the US or its allies...
Culture, Politics, War
God Urges Jews, Muslims to Stand their Ground in Dispute over Temple Mount
With tensions between Israelis and Palestinians on Jerusalem's Temple Mount reaching yet again another boiling point over the past year – as if 2020 hasn't been enough of a fail of a year – God once again urged both Jews and Muslims to resist pressure to compromise and said both religious groups should "fight to the last drop of blood"...
Entertainment, Politics, War
Iran Announces Soleimani Prize for Revolutionary Leaders Who Don't Die in a Drone Strike
Iranian President Hassan Rouhani announced this week the formation of a committee to award the ״Major General Soleimani Prize״. The prize is in memory of the infamous general who died last year from complications related to a hellfire missile and will be awarded for excellence in the category of "Struggle and Resistance". The award will be the centerpiece of an...
ISIS Announces 2021 Re-Branding: Islamic Safe Space; Berkeley Pledges Allegiance
ISIS has announced that, as part of its upcoming 2021 rebranding program, it will no longer be known as 'Islamic State' but would rather be known as 'Islamic Safe Space in Iraq and Syria'. The move is seen as an attempt to appeal more towards the younger generation and socially conscious millennials who have made just about everything you can...
Hamas Frustrated It Continues to Receive World Sympathy
Hamas has grown frustrated in recent months over its inability to convince the world that it is, indeed, a terrorist organization. "The Hamas brand just isn't sexy anymore," explained a Hamas member wearing an 'I Heart Terror' t-shirt. "Look at other successful terrorist organizations; all of them are unilaterally recognized as evil, terrible. In the name of Allah, what do...
Following SCOTUS Decision, Yemen tells Trump: 'Now You Know What Bombing Feels Like'
SUPREME COURT, USA and SANA'A, YEMEN — For years Yemen has been experiencing the (currently) worst humanitarian crisis of the 21st century, a country throat punched by war, famine, and pestilence, and much like Ivanka trying to mingle, they too are consistently ignored by global leaders. However, Yemeni diplomats felt a flicker of hope yesterday after watching the Supreme Court...
ISIS Practices Daily Affirmation: "Americans Are Still More Scared of Us Than COVID"
In a troubling week for members of the so-called 'Islamic State' – or whatever they call that last remaining mound of Syrian sand – the number of deaths from COVID-19 across the United States surpassed their wildest expectations of impact from any act of terror. Facing the reality that a disease was more effective than Jihad has created severe depression...
Hamas Environment Minister Vows to Cut Flag, Effigy Burning 20% by 2030
In one of the region's most ambitious initiatives to address climate change, Hamas Minister of the Environment Ahmet al-Buluti has vowed to reduce flag and effigy burning emissions 20% by 2030. "We must all do our part to confront the reality of global warming, and these targets will be an important step towards reducing our carbon footprint," al-Buluti said in...
ISIS, White Supremacists Announce Merger to Form Mega-Terror Group
In a move that will create a virtual monopoly on the terror industry, what remains of the radical Islamist group ISIS has agreed to merge with the worldwide white supremacist movement. The merger was announced in a joint press conference, covered by OAN news network, between ISIS leader Abu Ibrahim al-Qurashi and Richard Spencer, the president of the National Policy...
Iranian Nuclear Scientist Assassinated for Having 11 Dinner Guests
New York State Governor Andrew Cuomo has claimed responsibility for the assassination of top Iranian nuclear scientist Mohsen Fakhrizadeh, telling reporters he had no choice but to order the killing after he learned that Fakhrizadeh planned to host more than ten guests at a dinner party that evening. Cuomo said intelligence sources inside Iran witnessed the scientist pickup up a...


The Left Coaster: Hillary's Luck, and a Sunni Commitment?
Hillary's Luck, and a Sunni Commitment?
You have to wonder how giddy the Hillary camp is at poll numbers like these, where Donald Trump is pulling away from the GOP pack the harsher he sounds. We'll see how things stand after Trump takes incoming during tonight's debate.
Also, it is noteworthy that Saudi Arabia was able to assemble 34 nations in an anti-terrorism effort against Islamic extremism. It's good news that these are largely Sunni countries, who must now commit actual ground forces to the battle for this to have any real teeth. Now if the Saudis would only stop funding the extremism themselves, . . . .
Deacon Blues :: 5:48 PM :: Comments (1) :: TrackBack (0) :: Digg It!


The collection consists of photographs of soldiers of the 456 AAA Battalion, in uniform and casual dress, in Germany, France, Illinois, and Wisconsin (1942-1943). Also included is a letter from James S. Easley stationed with the 456th AAA Battalion in France, to his wife, in Mississippi (1945), and reunion newsletters of the 456th AAA Battalion (1982, 1984).
James S. Easley World War II photographs and letter, ms 2298, Hargrett Rare Book and Manuscript Library, The University of Georgia Libraries.


The overall purpose of trademark law is to prevent unfair competition by protecting the use of a symbol, word, logo, slogan, design, domain name, etc. that uniquely distinguishes the goods or services of a firm. This is to prevent confusion as to the source of goods or services. A copyright protects an original artistic or literary work fixed in a tangible medium of expression.
For a low cost, flat fee Perry Trademark Law provides clients with Trademark application services.
Perry Trademark Law conducts comprehensive screening and clearance searches of marks to determine their availability to clients. Search results are typically provided with a very short turnaround.
Perry Trademark Law is positioned to evaluate office actions issued by the USPTO and create a plan of action for response that best suits an entities needs.
Perry Trademark Law is positioned to monitor and maintenance an already issued mark by searching for potential infringing activities by others.
Perry Trademark Law provides all services related to required USPTO Renewal and Declaration of Continued Use, Sections 8 and 9 respectively.
Copyright laws protect original works of authorship fixed in a tangible medium of expression. Protected authorship includes literary, dramatic, musical and artistic works such as poetry, novels, movies, songs, computer software and architecture.
A trade secret is a formula, practice, process, design, instrument, pattern, commercial method, or compilation of information not general known or reasonably ascertainable by others by which a business can obtain an economic advantage over competitors or customers.


Who doesn't love getting mail? Now, children can role play mailing their letters at our Post Office playhouse with our interactive mailbox, sorting them in our old-fashioned pigeon-holed letter cabinet, and delivering the mail all around the village. That means fun no matter what role you play!


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Nintendo Playing Cards
Nintendo has finally returned to its roots of starting a life as a card maker with these Nintendo Playing Cards. One of the most influential video games of all times, this set features the Mario characters on your playing cards...
Weston Lanpher
Cool GadgetsGeeky
Nintendo has finally returned to its roots of starting a life as a card maker with these Nintendo Playing Cards. One of the most influential video games of all times, this set features the Mario characters on your playing cards. They are standard poker sized Mario and Zelda with the two joker cards having Bowser on it. If you miss bi pixels and have been frustrated with Playstation or CDs, here's your chance to live that era once again through your simple game of cards. Your Mario Bros game and the Nintendo NES nostalgia is now just a set of cards away.
Get it forGet it for $10.67
Gadget Flow Rating 9.4/10
Discovered on Feb 14, 2015, 9:00 am EST
Discovered by Weston Lanpher


Visitors who attended Fastener Fair Mexico 2019 are listed alphabetically below.
ALPHA STRUMBOS DE MÉXICO S. DE RL. DE CV.
AZTECA LIGHTING S.A. DE C.V.
CMP PRATSA S.A. DE C.V.
COMERCIAL VAZQUEZ MONTALVO, S.A. DE C.V.
COMERCIALIZADORA CHARLES S.A. DE C.V.
DISEKO SOLUCIONES, S.A. DE C.V.
DISTRIBUIDORA DE TORNILLOS MONJAC, S.A. DE C.V.
E&H PRECISION DE MEXICO S.A. DE C.V.
ELECTROPLATING MEXICO, S.A. DE C.V.
ELECTROTECNIA PLUS S.A.P.I. DE C.V.
EMPACADORA EL CENTENARIO S.A. DE C.V.
E-PLAST MX S.A DE C.V.
ESPECIALIDADES TERMICAS, S.A. DE C.V.
FIJACIONES ANTICORROSIVAS, S.A. DE C.V.
FLEXTRONICS MANUFACTURING AGUASCALIENTES S.A. DE C.V.
FORJADORA MEXICANA DE TORNILLOS, S.A. DE C.V.
GEOMEMBRANAS Y GEOSINTÉTICOS, S.A. DE C.V.
GEOMETRICA DE MEXICO S.A. DE C.V.
GRUPO COMERCIAL DITORBI, S.A. DE C.V.
GRUPO EMPAC SA. DE CV.
HERRAMIENTAS CASA MIRANDA S.A. DE C.V.
HIKER CARGO LOGISTICS S.A. DE C.V.
IC INTERNATIONAL COMMERCE LAWYERS & CONSULTANTS S.C.
IMPORTACIONES TOLEDO, S.A. DE C.V.
INDUSTRIA REAL, S.A. DE C.V.
INDUSTRIAS BUFAB DE MÉXICO, S.A. DE C.V.
JEDDA TTR, S. DE R.L. DE C.V.
LICON Y ASOCIADOS S.A. DE C.V.
LII UNITED PRODUCTS, S. DE R.L. DE C.V.
MARLLAM DE MATAMOROS, S.A. DE C.V.
METAL ONE DE MEXICO, S.A. DE C.V.
MEUSBURGER MEXICO S. DE R.L. DE C.V.
MT EQUIPOS Y ACCESORIOS INDUSTRIALES SA DE CV.
OEC FREIGHT S. DE R.L. DE C.V.
PAVCO MEXICO, S.A. DE C.V.
PEINES TANGENCIALES, S.A. DE C.V.
PEISA FASTENERS MEXICO, S.A. DE C.V.
PINTARRÓN ALFRA S.A. DE C.V.
POSCO MVWPC, S.A. DE C.V.
PRODUCTOS DE ALAMBRE SIMAR, S.A. DE C.V.
RENO DISTRIBUCIONES COMERCIALES S.A. DE C.V.
REPORTERO INDUSTRIAL MEXICANO S.A. DE C.V.
ROKI MEXICO S.A. DE C.V.
ROSCADOS Y PARTES INDUSTRIALES, S. A. DE C. V.
RPK MEXICO S.A. DE C.V.
SEW EURODRIVE MÉXICO, S.A. DE C.V.
SOMAKI DE MEXICO S.A.DE C.V.
TECHOS Y AISLAMIENTOS TERMICOS, S.A.S.
THREE BOND MANUFACTURING MEXICANA, S.A. DE C.V.
TOHKEN THERMO MEXICANA, S.A. DE C.V.
TORNILLOS ROU SAM SA. DE C.V.
TORNILLOS Y MUELLES SAN MARCOS, S.A. DE C.V.
TRAELO DE ASIA S. DE R. L. DE C. V.
TUBULARES MEXICANOS, S.A DE C.V.
UNICAR PLASTICS S.A. DE C.V.


Bacterial drug resistance is one of the most significant challenges to human health today. In particular, effective antibacterial agents against methicillin-resistant Staphylococcus aureus (MRSA) are urgently needed. A causal relationship between nasal commensal S. aureus and infection has been reported. Accordingly, elimination of nasal S. aureus reduces the risk of infection. Enzymes that degrade bacterial cell walls show promise as antibacterial agents. Bacteriophage-encoded bacterial cell wall-degrading enzymes exhibit intrinsic bactericidal activity. P128 is a chimeric protein that combines the lethal activity of the phage tail-associated muralytic enzyme of Phage K and the staphylococcal cell wall targeting-domain (SH3b) of lysostaphin.
Here we report results of in vitro studies evaluating the susceptibility of staphylococcal strains to this novel protein.
Using the broth microdilution method adapted for lysostaphin, we found that P128 is effective against S. aureus clinical strains including MRSA, methicillin-sensitive S. aureus (MSSA), and a mupirocin-resistant S. aureus. Minimum bactericidal concentrations and minimum inhibitory concentrations of P128 (1-64 μg/mL) were similar across the 32 S. aureus strains tested, demonstrating its bactericidal nature.
In time-kill assays, P128 reduced colony-forming units by 99.99% within 1 h and inhibited growth up to 24 h.
In an assay simulating topical application of P128 to skin or other biological surfaces, P128 hydrogel was efficacious when layered on cells seeded on solid media. P128 hydrogel was lethal to Staphylococci recovered from nares of healthy people and treated without any processing or culturing steps, indicating its in situ efficacy. This methodology used for in vitro assessment of P128 as an agent for eradicating nasal carriage is unique.
The novel chimeric protein P128 is a staphylococcal cell wall-degrading enzyme under development for clearance of S. aureus nasal colonization and MRSA infection. The protein is active against globally prevalent antibiotic-resistant clinical isolates and other clinically significant staphylococcal species including S. epidermidis. The P128 hydrogel formulation was bactericidal against Staphylococci including S. aureus recovered from the nares of 31 healthy people, demonstrating its in situ efficacy.
Antibiotic-resistant Staphylococcus aureus strains emerging from the community as well as hospital environments represent a global threat [1, 2], requiring new approaches to control this pathogen. The anterior nare is the major reservoir of S. aureus in humans; 80% of the human population may be carriers . A causal relationship between nasal colonization of S. aureus and serious infection has been established; thus, eliminating S. aureus nasal carriage may reduce the risk of infection [4, 5]. Coagulase-negative Staphylococci (CoNS) are known commensal flora of the skin and mucous membranes and also colonize human anterior nares. Recently CoNS have been recognised as opportunistic pathogens responsible for the increasing incidence of serious nosocomial infections, mainly because of their affinity for the foreign materials used in prosthetics and indwelling devices. Immunocompromised patients, including those undergoing haemodialysis, are especially susceptible to these infections [6, 7]. More than 80% of clinical CoNS strains and 30% to 40% of CoNS obtained from healthy carriers or patients from the community are resistant to methicillin .
Bactroban Nasal (Mupirocin ointment) has been approved for nasal clearance of S. aureus and significantly reduces the risk of postoperative staphylococcal infection in carriers . However, mupirocin resistance has already been reported, and its use is restricted in many countries. A superior product for intranasal prophylaxis in at-risk patients is therefore an unmet medical need. New chemical entities take longer to develop, and killing by broad-spectrum antibiotics is undesirable. Current efforts are therefore focused on pathogen-specific biological entities such as peptidoglycan hydrolases , antibodies , and other antimicrobial peptides and proteins . For example, lysostaphin is a bacterial peptidoglycan hydrolase that has been extensively studied for its antistaphylococcal activity in various animal models [13–15]. Bacteriophages are viruses that infect and kill bacteria and have co-evolved with bacterial defenses . Bacteriophages have been used for human therapy in several Eastern European countries for decades . Although they have not been used in clinical applications in Western countries, the United States Food and Drug Administration recently approved the use of bacteriophages to prevent bacterial contamination in meat . In addition, bacteriophages are a good source of cell wall-degrading enzymes, which have been evaluated as antibacterial agents [19–21].
P128 is a novel chimeric protein that derives its staphylococcal cell wall-degrading enzymatic domain from the gene product, ORF56, of bacteriophage K and the cell wall-targeting domain (SH3b) from Lysostaphin (Pubmed accession no. of Lysostaphin gene: M 15686.1). We have previously reported the construction of this novel chimeric protein and assignment of its peptidoglycan hydrolase activity to the Cysteine, Histidine-dependent AmidoHydrolase/peptidase (CHAP) domain. We also demonstrated the efficacy of P128 in nasal clearance of methicillin-resistant S. aureus (MRSA; strain USA300) in a rodent model . P128 is under development for topical indications including use against S. aureus nasal carriage. In this study we tested the antistaphylococcal activity of P128 by determining minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), time-kill kinetics, and activity against Staphylococci from human nares.
All S. aureus strains used in the study are listed in Table 1. These include 30 clinical strains (27 MRSA strains and 3 MSSA strains) from the Public Health Research Institute, New Jersey and two USA 500 strains.
MIC was determined using a modification of the CLSI broth microdilution method. P128 was tested at 256 to 0.125 μg/mL. S. aureus ATCC 25923 and S. carnosus ATCC 51365 were used as control strains. MBC was determined following the CLSI procedure by plating 100 μL from the MIC, MIC × 2, MIC × 4, and MIC × 8 wells on LB agar, and incubating the plates overnight at 37°C. Strains 1-30 constitute a global panel of distinct clinical isolates (MRSA, strains1-27; MSSA, strains 28-30) obtained from the Public Health Research Institute (NJ, USA); strains 31 and 32 are USA500.
P128 protein was cloned and expressed under the inducible T7 expression system in E. coli ER2566 strain. Details of cloning and design of the P128 clone-construct were reported previously (22).
To generate a purified preparation of P128 for the studies reported in this work, expression of P128 protein in E. coli ER2566 was induced with 1 mM IPTG, at 37°C for 4 h. The induced cell pellet was lysed and the protein in the supernatant was subjected to 0-50% ammonium sulphate precipitation using solid ammonium sulphate at 4°C. The precipitate was dialysed against 25 mM Tris HCl buffer pH 8.0, passed through an anion exchange column. The unbound fraction (flow through), containing P128 protein, was bound to a cation exchange column using 50 mM sodium acetate buffer at pH 6.0. The bound protein was eluted using a linear gradient of 0 to 0.5 M sodium chloride. Fractions containing P128 protein were extensively dialysed against saline and used for all the studies.
The MIC was determined using a modified Clinical and Laboratory Standards Institute (CLSI) broth microdilution procedure . Briefly, microtiter wells were pre-coated with 0.5% bovine serum albumin (BSA) to prevent nonspecific P128 adherence to the polystyrene plate, based on the method published for lysostaphin . Two-fold dilutions of P128 were prepared in Mueller Hinton broth (MHB; Himedia) supplemented with 0.1% BSA (Sigma Aldrich), and 50 μL aliquots of the P128 dilutions (0.125-256 μg/mL) were added to the wells. Bacterial suspensions (0.5 McFarland standard) were diluted in MHB to achieve 1 × 106 colony-forming units (CFU) per mL. Then 50 μL aliquots of the cell suspension were added to wells containing P128. Plates were incubated under static conditions at 35°C for 18 h. The MIC was defined as the lowest concentration of P128 in which no visible growth was observed at the end of the incubation period. The MBC was also determined using the CLSI procedure. Briefly, 100 μL from the MIC, two times MIC (MIC × 2), four times MIC (MIC × 4), and eight times MIC (MIC × 8) wells were plated on Luria Bertani (LB) agar and incubated at 37°C overnight.
MIC of Vancomycin was determined for a panel of S. aureus isolates that represented the MIC range of P128 (1-64 μg/mL) using the CLSI broth microdilution method. Vancomycin was tested at concentrations of 0.125-256 μg/mL, and MICs were read manually after 24 h of incubation. MBC was also determined using the CLSI procedure. The reference strain, S. aureus ATCC 25923 was used for quality control of the assay, in case of both P128 and Vancomycin MIC and MBC determinations.
The kinetics of P128 bactericidal activity were assessed in vitro using six S. aureus strains: BK#13237, BK#9894, BK#14780, BK#8374, BK#9918, and BK#19069. The cryopreserved test strains were plated on LB agar plate and incubated overnight at 37°C. Several well-isolated colonies were picked up and suspended in MHB broth; the turbidity was then adjusted to 0.5 McFarland standard (about 108 CFU/mL). The initial inoculum was prepared by inoculating 10 μL of each test bacterial suspension into 20 mL MHB supplemented with 0.1% BSA. After 1 h in a shaker incubator (37°C, 200 rpm), 2.7 mL aliquots of the culture were dispensed into four tubes, and 0.3 mL P128 was added. A 0.3 mL aliquot was immediately removed to determine the initial CFU (0 h). Incubation was continued, and 0.3 mL aliquots were taken at 1, 2, 4, 8, and 24 h. The cultures were serially diluted in sterile saline immediately after sampling and plated on MHB agar. After overnight incubation of the plates, CFU were determined. The time-kill curve was plotted based on bacterial survival at the sampling intervals .
P128 hydrogel was formulated with hydroxyethyl cellulose (0.42%), propylene glycol (0.75%), and glycerin (2.25%) as the main excipients along with P128 protein. A formulation that contained physiological saline in place of P128 (referred to as buffer gel) served as a negative control. LB agar was poured into 24-well tissue culture plates (Tarson). S. aureus (BK#13237) cells at 103 CFU/well (Figure 1) and 102 CFU/well (Figure 1) were seeded on LB agar in the microwells. P128 gel was diluted two-fold in buffer gel to contain P128 protein at a concentration range of 100 to 1.56 μg/mL. P128 gel preparations were applied to wells and the plates were incubated at 37°C for 18 h. At the end of incubation, 20 μL iodonitrotetrazolium chloride (INT dye; Loba Chemie) prepared in 50 mM sodium phosphate buffer, pH 7.0 (30 mg/mL) was added to the wells to visualize viable cells.
Efficacy of P128 gel formulation applied to S. aureus on agar surface. A hydrogel formulation containing P128 protein (100 to 1.56 μg/mL) was tested for bactericidal activity when applied topically on S. aureus strain BK#13237 cultured on LB agar: (a) 103 CFU/well, (b) 102 CFU/well. Well #1 represents the media control, and well #2 represents the cell control. In both (a) and (b), P128 gel preparations (100-1.56 μg/mL) were added to wells #3-9; P128 protein formulated in physiological saline (100 μg/mL) was added in well #10 as a positive control; buffer gel was added to well #11 as a negative control. INT dye was added to the visualize growth of the surviving bacteria.
Activity of P128 was tested in a buffer that simulated the ionic composition of nasal fluid. The simulated nasal fluid (SNF) contained 0.87% NaCl, 0.088% CaCl2. 2H20, 0.31% KCl, and 0.636% BSA . The S. aureus COL strain was subcultured in LB medium from an overnight culture and grown at 37°C and 200 rpm until the OD600 reached 1.0 to 1.5 (5 × 108 CFU/mL). 100 μL of this cell suspension (5 × 107 CFU) was centrifuged at 3000 × g for 10 min and the cell pellet was suspended in 100 μL of SNF. 100 μL of P128 prepared in SNF (1.5 μg/mL) was added to the cells. As a positive control, P128 contained in physiological saline was added to cells suspended in physiological saline. After addition of P128, tubes were incubated for 1 h in a shaker incubator at 37°C, 200 rpm. Cells were then pelleted and resuspended in 1 mL LB, and 10-fold dilutions were plated on LB agar and incubated at 37°C overnight. Cells treated with SNF or saline served as untreated cell controls.
Nasal commensal Staphylococci of 31 healthy people were characterized and evaluated for sensitivity to P128. A dry swab (Copan Diagnostics) was inserted into each nostril, rotated six times to cover the entire mucosal surface of the anterior nare, and slowly withdrawn. The swab from one nostril of each individual was immersed in a vial containing 200 μL P128 hydrogel (40 μg/200 μL), and a swab from the other nostril was immersed in a vial containing 200 μL buffer gel (control). The vials were placed in a biosafety cabinet for 1 h at ambient temperature (about 25°C). The entire vial contents were then spread on blood agar plates and incubated overnight at 37°C. CFUs recovered were characterized in terms of colony morphology, hemolysis on blood agar, Gram stain, and a HiStaph identification kit (Himedia).
P128 is a bacteriophage derived staphylococcal cell-wall degrading enzyme. This protein is under development in our laboratory for topical therapeutic use in humans. In this study, we tested the bactericidal activity of P128 protein on globally prevalent S. aureus clinical strains. We assessed the biological activity of P128 using various in vitro assays and under conditions designed to simulate physiological conditions.
P128 protein preparations used in this study were of > 95% purity. The protein expressed was in the soluble form in a standard E. coli expression system and purified using a 2-step ion-exchange chromatography procedure .
Determination of MIC and MBC is a commonly used method to assess susceptibility to antimicrobial agents. We determined the MIC and MBC of P128 for a panel of 31 globally represented strains of S. aureus using modified CLSI methods . Microtiter plate wells were pre-coated with BSA before adding P128 to minimize nonspecific adherence and loss of protein to the polypropylene surface. The MIC of P128 for the various strains of S. aureus ranged from 1 to 64 μg/mL (Table 1). The MIC at which 50% of the strains tested were inhibited (MIC50) was 8 μg/mL. The MBC of P128 across S. aureus strains tested also ranged from 1 to 64 μg/mL; and the MBC50 was found to be 16 μg/mL (Table 1).
MIC and MBC of Vancomycin were determined using the same procedure that was used in case of P128. For the reference strain, S. aureus ATCC 25923 MIC and MBC of Vancomycin was found be 0.5 μg/mL and 2 μg/mL respectively. These values correlate with the reported MIC and MBC of Vancomycin for this strain, validating the assay used in this work. Vancomycin was also tested on a panel of S. aureus strains that represented the MIC range of P128 (1 to 64 μg/mL). MIC of Vancomycin for these strains ranged from 0.5 to 1 μg/mL and MBC ranged from 1 to 4 μg/mL (Table 2).
MIC was determined by modified broth microdilution method following the CLSI procedure. Vancomycin test concentration was in the range of 256 to 0.125 μg/mL. S. aureus ATCC 25923 was used as the control strain. MBC was determined following the CLSI procedure by plating 100 μL from the MIC, MIC × 2, MIC × 4 and MIC × 8 wells on LB agar and incubating the plates at 37°C overnight. The strains used here span the MIC range of P128.
Since MIC relates to growth inhibition activity of an antimicrobial agent, MBC may be a more appropriate measure of activity of P128 which is bactericidal in action.
Time-kill assays were performed in accordance with the CLSI guidelines, with a starting inoculum of 5 × 104 CFU/mL and, various multiples of the MICs. The objective of this assay was to evaluate concentration-dependent bactericidal activity. In order to find the optimal concentration required to achieve and maintain > 99.99% killing upto 24 h, sub-MIC levels were not considered. The detection limit of the time kill curve was 10 CFU/mL.
We determined the number of viable S. aureus cells remaining at different time intervals after adding P128 protein. Figure 2 shows the time-kill curves of P128 for six representative strains of S. aureus, which included five MRSA strains and one MSSA strain. P128 showed rapid, dose-dependent bactericidal activity against the MSSA and MRSA strains tested, killing of 99.99% of cells in all six strains tested within 1 h at the respective MIC concentration. At the MIC, growth was inhibited up to 24 h for all five MRSA strains and up to 8 h for the MSSA strain (BK#9918). However, the cells of BK#9918 that grew after 8 h were susceptible to P128 (data not shown). Since a concentration 4× the MIC inhibited growth of this strain for up to 24 h, we surmised that higher concentrations of P128 or repeated treatments may be required in such cases.
Kill-kinetics of P128 on S. aureus strains. Time-kill curves of P128 at three different concentrations (MIC, MIC × 4, and MIC × 16) on five MRSA and one MSSA strains are shown. Cell control was maintained simultaneously for each strain.
The efficacy of P128 hydrogel was tested on solid culture medium to simulate the conditions of topical nasal application.
The assay format was designed to check availability of the protein when applied as a gel formulation. The objective was also to test efficacy of P128 gel applied to a surface where low numbers of bacterial cells are present. We have used a range of 100-1 μg/mL of protein concentration in the gel formulation. P128 gel showed complete clearance at concentrations up to 1.56 μg/mL (Figure 1).
Functional efficiency and structural stability of enzymes can generally be influenced by pH, temperature, and the composition and concentrations of metal or inorganic ions in the reaction milieu. Our primary concern was that monovalent and divalent ions present in nasal fluid may have a deleterious effect on P128 activity. We therefore evaluated the activity of P128 in a composition that simulated the ionic content of normal human nasal fluid. We found that P128 reduced the staphylococcal viable count (CFU) by five orders of magnitude in SNF, comparable to the activity observed in case of P128 in physiological saline. Cells incubated in SNF that did not contain P128 were unaffected (Figure 3). These results indicate that the protein would not be influenced by the ionic content of human nasal fluid.
P128 activity in simulated nasal fluid. Bactericidal activity of P128 against S. aureus strain COL was tested under conditions simulating the ionic composition of human nasal fluid.
Secreted products and components such as exotoxins, exoenzymes, surface-associated adhesins, and capsular polysaccharide play a role modulating host responses to S. aureus infection . Production of capsular polysaccharide type 5 by Staphylococci has been reported in a study using a mouse model of S. aureus nasal colonization . The same study also showed the inability of a capsule-defective mutant to persist in mouse nares, indicating that S. aureus is encapsulated in the nares. The rate of methicillin resistance among CoNS isolates colonizing anterior nares of patients undergoing haemodialysis is reported to be higher than that of S. aureus isolates; this is accompanied by the lack of susceptibility to other classes of antibiotics . Although S. epidermidis is responsible for most CoNS infections, other CoNS species have been associated with a variety of human diseases . For example, S. haemolyticus is the second most commonly encountered species in clinical infections, and S. lugdunensis is a more recently described CoNS species .
In this context, we evaluated the bactericidal activity of P128 on S. aureus and other staphylococcal species recovered from human nares. As the first step, we characterized the nasal commensal bacteria of 31 healthy people. Speciation was carried out using the HiStaph identification kit and the S. aureus carriage rate was also determined. Nasal Staphylococci of 71% of the healthy people sampled consisted of CoNS species, predominantly S. epidermidis and S. aureus was found in the remaining 29% of people. Other CoNS among nasal commensal bacteria included S. haemolyticus and S. lugdunensis (Table 3). We examined nasal commensal populations in two randomly selected healthy people for comparability between the two nares with respect to bacterial load and staphylococcal species present and found both nares to be comparable (data not shown).
We then evaluated the activity of P128 hydrogel on nasal Staphylococci of 31 healthy people.
In case of nasal swabs immersed in buffer-gel, colonies were numerous, ranging from 103 - 105 CFU; estimated based on results of a preliminary experiment, where S. aureus cells of known CFU counts (103, 104 and 105 CFU) were plated to vizualize the pattern of growth after overnight incubation of plates (data not shown). Of the swabs immersed in P128 hydrogel, 4/31 showed > 99.99% reduction in staphylococcal cell counts, 17/31 showed 99.9% reduction, 5/31 showed 99% reduction, and 5/31 showed 90% reduction (Table 4). A few colonies that grew on the plate containing P128 were found to be sensitive to the protein when tested, and hence apparently escaped the protein action.
This finding shows that P128 is bactericidal to nasal staphylococcal isolates. However, we did not evaluate the presence of capsular polysaccharides, which may be assessed in future studies in our laboratory. It is important to note that the cells were treated with P128 hydrogel immediately after isolation (i.e., without exposure to any other medium or subjection to any steps of cultivation). We conclude that both S. aureus and CoNS are susceptible to P128 in the physiological state relevant to nasal carriage. Considering the pathogenic potential and multidrug resistance of these species, it is significant that these species were fully sensitive to P128. Further studies are needed to determine the MIC and MBC of P128 on CoNS.
Reports point to the endogenous origin of most infective S. aureus isolates and MRSA carriage poses an increased risk for invasive infections compared with MSSA carriage [30, 31]. The worldwide spread of MRSA strains, which are often multidrug-resistant , combined with limited therapeutic options necessitates new approaches to combat this pathogen. Recent findings emphasize that commensal CoNS strains are also potential threats . Therefore an antibacterial agent, exemplified by P128, which can target antibiotic resistant S. aureus as well as other clinically significant Staphylococci would meet the current medical need and warrants further development.
This report describes the development and in vitro biological characterization of a chimeric antistaphylococcal protein designated P128, which exhibits rapid and selective antibacterial activity at low MIC values against a broad range of staphylococcal species, including numerous clinically relevant S. aureus strains.
The MIC and MBC of P128 on a global panel of clinical isolates ranged from 0.5 to 64 μg/mL. P128 showed in vitro microbicidal activity against a wide variety of staphylococcal species, including many clinically relevant S. aureus strains. With an MBC50 of 16 μg/mL, the protein was bactericidal against every S. aureus strain tested. P128 time-kill kinetics were determined at MIC and higher concentrations on select isolates, and P128 was found to rapidly reduce cell numbers by 99.99%. To develop P128 as a treatment to eliminate human nasal carriage, P128 was formulated as a hydrogel and tested on nasal Staphylococci recovered from healthy people. The protein was able to kill S. aureus under conditions representing physiological conditions. Taken together, our findings demonstrate that P128 exhibits excellent antistaphylococcal properties and warrants development for therapeutic use.
The authors thank Dr. J Ramachandran for his support, review of data and key suggestions in this work. The authors would like to acknowledge the scientific staff at Gangagen, whose help and cooperation aided in the completion of this work. The authors thank Dr. Barry Kreiswirth, PHRI, New Jersey for providing global panel of S. aureus isolates and Dr. M. Jayasheela for reviewing the manuscript.
BS and AV participated in the study design and coordination and data interpretation. AV, SD, PR and RP evaluated the efficacy of P128 gel in nasal Staphylococci experiments. JR, RP, PR, SD, and NN performed P128 MIC and MBC assays. JR and PR performed time-kill curve experiment. VP tested P128 activity in SNF, and RC evaluated the efficacy of P128 hydrogel in the agar surface assays. AV also helped draft the manuscript. All authors read and approved the final manuscript.


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Medina County Criminal Defense Lawyers for Repeat Violent Offenders
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Repeat offenses occur when an individual commits a crime for which they have previously been convicted. Convictions that qualify as prior offenses in Ohio are defined under state law and usually must take place within a specified period of time from the most recently committed offense. For example, a conviction for driving under the influence of alcohol or drugs is considered to be a repeat offense if the offender has a prior conviction for DUI within six years.
Most categories of crimes increase the severity of the offense from a misdemeanor to a felony and increase the penalties imposed if you are convicted including fraud, drug and alcohol-related driving offenses, or possession and sale of drugs. Even minor offenses can be elevated to the misdemeanor level if the offender has a prior criminal record.
Repeat Violent Offenders
Designating an individual as a repeat violent offender begins when prosecutors present the case to a grand jury for an indictment or file an information charging an offender with committing a crime. At the arraignment, a defendant is entitled to receive notice that the prosecution intends to offer into evidence a certified copy of the official record of prior convictions of the accused. Defendants provided with notice of the prior convictions must give notice to the prosecution of an intention to challenge or object to the record of convictions.
Examples of Repeat Offense Laws in Ohio
Violent offenders or those charged with violations of the drug laws who have prior convictions for the same offenses are at risk of a judge adding an additional 10 years to their sentences upon conviction. A person charged with a felony who uses, possesses, or brandishes a firearm during the criminal act can receive an additional three years added to the sentence at the discretion of the judge.
Repeat DUI Offenses in Ohio
The biggest impact of repeat offenses on the largest group of people might be in cases involving driving under the influence of alcohol or drugs. A first offense DUI is usually charged as a misdemeanor and punished with a fine ranging from $375 to $1075, a three-day minimum jail sentence or three-day driver intervention program, and a license suspension of six months to three years.
A person charged with DUI who was previously convicted of DUI within six years faces penalties of a fine ranging from $525 to $1,625, a minimum jail sentence of 20 days, and a license suspension ranging from one year to five years. If the accused has two prior convictions within six years, the penalties increase to a fine up to $2,750 with a minimum of $850, at least 30 days in jail, and a loss of driving privileges for up to 10 years.
A fourth or fifth DUI conviction within six years is elevated to a felony charge with a minimum incarceration period of 60 days and a maximum sentence of five years in prison. Fines range from $1,350 to $10,500, and license suspension can be for the rest of a person's life. If a person is charged with committing a DUI for the sixth time, the fine can be as high as $10,500, and incarceration is a minimum of 120 days up to a maximum of five years. The look-back period is extended from six years to 20 years. This means that convictions as far back as 20 years ago can be considered in charging the individual as a repeat offender.
Sentencing Discretion and Repeat Offenses
Judges are given a great deal of discretion in the sentences they impose. State sentencing guidelines suggest factors judges should consider when deciding the type and severity of the punishment for a person convicted of committing a crime.
One factor judges take into consideration is the prior criminal record of the offender. Particular attention is given to the type of prior offenses. An offender with a history of committing the same type of offenses can expect to receive a harsher sentence.
If you are a repeat offender, you need qualified, experienced defense attorneys on your side. The lawyers at Erb Legal LLC will fight for you. We will examine every angle to possibly have your prior record removed from consideration, reduce your charges, or limit the penalties. Contact us at 330-368-4499 for a free, initial consultation. We serve Medina County, Summit County, and Wayne County, Ohio.
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