This entry was posted in Non classé and tagged 2004, 3, cats, Frimousse, November, Tigri on 15 March 2017 by Le Maitre de Frimousse.
This entry was posted in Non classé and tagged 2004, rember, septembre on 10 March 2017 by Le Maitre de Frimousse.
Alone, in his garden, on August 23rd 2004.
With the "team", in our house, on May 22nd 2004.
This entry was posted in Non classé and tagged 2004, Frimousse, garden, home, RouXy, Tigri on 8 March 2017 by Le Maitre de Frimousse.
RouXy and Jump on December 30th, 2003. Jump died in February 2007. He was only 12.
Tigri, RouXy, and Frimousse on March 18, 2004. Tigri died in September 2012. He was 9.
This entry was posted in Non classé and tagged 2003, 2004, died, Frimousse, Jump, RouXy, Tigri on 6 March 2017 by Le Maitre de Frimousse.


text:
<?xml version="1.0" encoding="UTF-8"?>

<!--
Copyright 2014-2022 Grzegorz Slowikowski (gslowikowski at gmail dot com)

Licensed under the Apache License, Version 2.0 (the "License");
you may not use this file except in compliance with the License.
You may obtain a copy of the License at

  http://www.apache.org/licenses/LICENSE-2.0

Unless required by applicable law or agreed to in writing,
software distributed under the License is distributed on an
"AS IS" BASIS, WITHOUT WARRANTIES OR CONDITIONS OF ANY
KIND, either express or implied.  See the License for the
specific language governing permissions and limitations
under the License.
-->

<project xmlns="http://maven.apache.org/POM/4.0.0" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:schemaLocation="http://maven.apache.org/POM/4.0.0 http://maven.apache.org/xsd/maven-4.0.0.xsd">

    <modelVersion>4.0.0</modelVersion>
    <groupId>org.scoverage.scoverage-maven-samples.different-test-frameworks</groupId>
    <artifactId>scecs2</artifactId>
    <version>1.4.12-SNAPSHOT</version>
    <packaging>jar</packaging>

    <name>SCoverage Maven Samples : Different Test Frameworks : Specs2</name>

    <properties>
        <project.build.sourceEncoding>UTF-8</project.build.sourceEncoding>
        <project.reporting.outputEncoding>UTF-8</project.reporting.outputEncoding>

        <scala.version>2.11.12</scala.version>
        <sbt.version>0.13</sbt.version>

        <specs2.version>2.3.13</specs2.version>

        <compiler.plugin.version>3.8.0</compiler.plugin.version>
        <project-info-reports.plugin.version>3.0.0</project-info-reports.plugin.version>
        <site.plugin.version>3.7.1</site.plugin.version>
        <surefire.plugin.version>2.22.1</surefire.plugin.version>

        <sbt-compiler.plugin.version>1.0.0</sbt-compiler.plugin.version>
        <specs2.plugin.version>0.4.4</specs2.plugin.version>

        <scoverage.plugin.version>1.4.11</scoverage.plugin.version>
    </properties>

    <dependencies>
        <dependency>
            <groupId>org.scala-lang</groupId>
            <artifactId>scala-library</artifactId>
            <version>${scala.version}</version>
        </dependency>

        <dependency>
            <groupId>org.specs2</groupId>
            <artifactId>specs2_2.11</artifactId>
            <version>${specs2.version}</version>
            <scope>test</scope>
        </dependency>
    </dependencies>

    <build>
        <sourceDirectory>${basedir}/src/main/scala</sourceDirectory>
        <testSourceDirectory>${basedir}/src/test/scala</testSourceDirectory>

        <plugins>
            <plugin>
                <groupId>org.apache.maven.plugins</groupId>
                <artifactId>maven-compiler-plugin</artifactId>
                <version>${compiler.plugin.version}</version>
                <configuration>
                    <skipMain>true</skipMain>
                    <skip>true</skip>
                </configuration>
            </plugin>

            <plugin>
                <groupId>com.google.code.sbt-compiler-maven-plugin</groupId>
                <artifactId>sbt-compiler-maven-plugin</artifactId>
                <version>${sbt-compiler.plugin.version}</version>
                <executions>
                    <execution>
                        <goals>
                            <goal>compile</goal>
                            <goal>testCompile</goal>
                        </goals>
                    </execution>
                </executions>
            </plugin>

            <plugin>
                <groupId>org.apache.maven.plugins</groupId>
                <artifactId>maven-surefire-plugin</artifactId>
                <version>${surefire.plugin.version}</version>
                <configuration>
                    <skipTests>true</skipTests>
                </configuration>
            </plugin>

            <plugin>
                <groupId>com.mmakowski</groupId>
                <artifactId>specs2-maven-plugin</artifactId>
                <version>${specs2.plugin.version}</version>
                <!-- Override dependencies in order to avoid an error in https://repo.maven.apache.org/maven2/com/github/scala-incubator/io/scala-io-file_2.11/0.4.3/scala-io-file_2.11-0.4.3.pom -->
                <!-- (org.scala-lang.modules:scala-parser-combinators_2.11 depenedncy defined twice with different versions, version 1.0.0 does not exist in the repository) -->
                <dependencies>
                    <dependency>
                        <groupId>com.github.scala-incubator.io</groupId>
                        <artifactId>scala-io-core_2.11</artifactId>
                        <version>0.4.3-1</version>
                    </dependency>
                    <dependency>
                        <groupId>com.github.scala-incubator.io</groupId>
                        <artifactId>scala-io-file_2.11</artifactId>
                        <version>0.4.3-1</version>
                    </dependency>
                </dependencies>
                <executions>
                    <execution>
                        <id>test</id>
                        <phase>test</phase>
                        <goals>
                            <goal>run-specs</goal>
                        </goals>
                    </execution>
                </executions>
            </plugin>

            <plugin>
                <groupId>org.scoverage</groupId>
                <artifactId>scoverage-maven-plugin</artifactId>
                <version>${scoverage.plugin.version}</version>
            </plugin>
        </plugins>

        <pluginManagement>
            <plugins>
                <plugin>
                    <groupId>org.apache.maven.plugins</groupId>
                    <artifactId>maven-site-plugin</artifactId>
                    <version>${site.plugin.version}</version>
                </plugin>
            </plugins>
        </pluginManagement>
    </build>

    <reporting>
        <plugins>
            <plugin>
                <groupId>org.apache.maven.plugins</groupId>
                <artifactId>maven-project-info-reports-plugin</artifactId>
                <version>${project-info-reports.plugin.version}</version>
                <reportSets>
                    <reportSet>
                        <reports>
                            <report>index</report>
                        </reports>
                    </reportSet>
                </reportSets>
            </plugin>

            <plugin>
                <groupId>org.scoverage</groupId>
                <artifactId>scoverage-maven-plugin</artifactId>
                <version>${scoverage.plugin.version}</version>
                <reportSets>
                    <reportSet>
                        <reports>
                            <report>report</report> <!-- select only one report from: report, integration-report and report-only reporters -->
                        </reports>
                    </reportSet>
                </reportSets>
            </plugin>
        </plugins>
    </reporting>

</project>

redpajama_set_name: RedPajamaGithub


This is a placeholder page for Larry Wholmuth, which means this person is not currently on this site. We do suggest using the tools below to find Larry Wholmuth.
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text:
Q: Chat Button Display During Hours of Operation I am trying to get a chat button to display during hours of operation and then a closed statement to display after hours.
I am able to get it to display the chat button only but it doesn't go away after hours.
Hours of operation are 6:30AM - 3:30PM Central Time
Thanks in advance!
  var openingTime = new Date();
  openingTime.getUTCHours(11); openingTime.getUTCMinutes(30);

  var closingTime = new Date();
  closingTime.getUTCHours(20); closingTime.getUTCMinutes(30);

var d = new Date();

if(((d.getUTCHours() >= openingTime.getUTCHours() && d.getUTCHours() <= closingTime.getUTCHours() ) && (d.getUTCDay() >= 1 && d.getUTCDay() <= 5) )) {
    $(".open").show();
    $(".closed").hide();
}
else {
     $(".closed").show();
    $(".open").hide();
}

A: From the code, I see that you are using getUTCHours and getUTCMinutes to set the hour and minute. However these two methods are only for getting the hour and minute from the Date object.
Instead use setUTCHours and setUTCMinutes to set the hour and minute
This might not be the root of the problem, but it's a small error I found when reading the code.
Hope it helps.

redpajama_set_name: RedPajamaStackExchange


text:
About VeriDaaS
Aviation Fleet
Geiger Mode LiDAR
Geospatial Solutions
VeriDaaS Data Library
5G Telecom
Aviation TOD
Ecology & Conservation
Obstacle and terrain data meeting the data quality requirements for in-flight and ground-based applications provide significant safety benefits for international civil aviation. The International Civil Aviation Organization (ICAO) requires all ICAO-participating States to ensure the availability of digital terrain and obstacle data for four areas covering the territory of the State as well as well as the operationally significant areas around an airport.
The terrain and obstacle data collected must comply with the regulatory requirements defined for each area. The obstacle and terrain data is stored digitally and made available as digital data sets for operational use by airspace users and air navigation service providers The four basic coverage areas are below:
The entire territory of the State
Constituted by 2d, a 45-km radius section around the airport (terminal control area), and by three other sections: 2a, 2b, and 2c.
Aerodrome or heliport area extending from the runway center line to 90 m and from other parts of the aerodrome movement area to 50 m.
The radio altimeter operating area runway, category II or III; the width of Area 4 is 60 m on either side of the extended runway center line, while the length is 900 m from the runway threshold.
VeriDaaS' TOD service provides airports, airlines, and air-traffic authorities with cost-effective digital data to help facilitate ICAO compliance, as well as for other applications in the aviation domain. Our datasets can help relevant authorities in the State meet the data quality requirements specified by ICAO as well as the national requirements defined by the State (i.e., one collection can satisfy multiple uses). Our data service is designed for cost-effective collection and provision of required data and contributes to decreased costs for aviation by providing the tools needed to identify possibilities to reduce fuel waste, cut carbon emissions, augment cargo loads and support the implementation of Unmanned Aircraft Systems Traffic Management(UTM).
WHY VERIDAAS?
Data compliant with ICAO requirements and industry standards as prescribed by ISO, RTCA (DO) and EUROCAE (ED). We can help simplify your ICAO Annex 15 TOD compliance by remaining current and up to date on all current standards and future amendments.
Multi-Disciplinary Skillset
Our highly skilled teams include aviation and geospatial experts with extensive experience across aeronautical and terrestrial surveying, data processing and software development. Our teams operate within discrete workflows according to primary areas of expertise, guided by well-defined and documented rules, standards, and processes.
An Innovative and Economical Solution
VeriDaaS uses the most technically innovative geospatial technology available to deliver the highest-quality TOD solution possible. The efficiency of our Geiger-mode aerial LiDAR helps to make ICAO compliance more cost effective than conventional terrestrial surveys, and we provide easy access to seamless data of homogenous terrain and obstacle data that is centrally located.
Access to Proprietary Geiger-mode LiDAR
Our proprietary high-density Geiger-mode LiDAR systems, developed by L3-Harris, collect high-resolution data of up to 50 pts per m2 providing a singular homogenous data source across all areas. The superior accuracy of Geiger-mode data (less than 10 cm vertical accuracy) enables optimization of flight plans and improved utilization of airspace.
Geiger-mode LiDAR collection is three to four times faster than traditional methods. This enables data to be collected in a shorter timeframe, reducing both overall cost and disruption to airspace. In addition, data can be collected from a higher altitude and at night, further reducing airspace disruption.
Geiger-mode LiDAR detects smaller features, such as antennas, which are often missed by other technologies. Our high-resolution datasets also meet the needs of many other use cases, providing additional economic value beyond TOD requirements. For example, our datasets can support the development of emerging aviation activities such as local unmanned commercial drone deliveries and urban air taxis.
VeriDaaS and its partners provide multiple avenues of access to LiDAR point clouds and TOD-specific data layers. Moreover, our sophisticated change-detection algorithms utilize satellite imagery to identify terrain and obstacle changes, triggering alerts when data updates are required.
Example Imagery
Click on image to expand.
Email: info@veridaas.com
VeriDaaS is a Denver-based leading geospatial solutions provider focused on providing data, products, and solutions, based on the best data available, at a national level.
© Copyright VeriDaaS Corporation. All Rights Reserved.

redpajama_set_name: RedPajamaCommonCrawl


To give you an idea of how Ultsearch uses the PPC engines in his grand scheme, consider an example.
Overture -- A PPC company. Furnishes search results for Ultsearch's site.
In this scenario, let's suppose that XYZ.com would like to rank first among the search results for the search term 'casino' in all sponsored search matches.
XYZ.com goes to Findwhat.com, creates a new account, and then proceeds to bid, for example $3.50, for each successful click to its site from any PPC result sponsored by Findwhat.
This is where Ultsearch comes in. Ultsearch has worked out a partnership/affiliate relationship with Findwhat through which he receives a cut of the bid amount generated from each successful clickthrough to XYZ.com that originates from one of his Websites.
And Bob, who likes casinos, happens upon one of Ultsearch's many cookie-cutter Websites. He notices that there is a link for casinos, so he clicks that link, and is taken to a list of sponsored results for casino sites. Noticing XYZ.com's top ranked listing, he clicks the sponsored link, and proceeds to XYZ.com for casino fun and excitement.
In this example, XYZ.com pays Findwhat $3.50 for referring Bob, as this was the amount it bid per click. Findwhat then pays Ultsearch a cut (30-50%), as the referral originated on one of his sites. Net take for Ultsearch: $1.75 for one click.
So it follows that, if you run Websites that receive lots of traffic, you can partner with PPC search engines like Findwhat to split the revenues generated by each click to a sponsored listing that originates on one of your sites. The revenue per click will vary depending on the search term, from just 1¢ to over $20 per click.
As an example, the term 'casino' currently has a maximum advertiser bid of $30 per click by Royal Casino! In contrast, a less commercial term like 'paintings' has a maximum advertiser bid of only $0.33 per click.
Now this is all fine and dandy and thousands of individuals on the 'net now affiliate with PPC engines in one capacity or another to boost the potential revenues they can achieve through their Websites. Yet we've only barely scratched the surface on how Ultsearch operates.
A Real World Example - Could This Be You?
To get some background on the way Ultsearch conducts business, let's take a look at the story of The Underdogs, a very popular Website that's home to one of the largest collections of abandonware and shareware games on the Web. In March of 2002, Sarinee, owner of The Underdogs, neglectfully forgot to renew his domain, as he didn't receive the warning emails his registrar had sent him about the looming expiry of his domain.
Unbeknownst to Sarinee, his expired domain was quickly grabbed the moment it became available -- by Ultsearch.
Now, although the domain name changed hands, the site was still heavily trafficked-due in large part to a network of reciprocal link partners and high search engine rankings that Sarinee had established over the years. The Underdogs was still receiving hundreds of thousands of visitors a day, even though the old Website was no longer active. Ultsearch quickly converted the thriving shareware site into his facsimile PPC directory to make money off the established traffic that was still being channelled to the site.
As a side note, many well-known companies have sued Ultsearch over this practice and failed, because what he does is still considered a "legitimate business" under WIPO arbitration rules).
Each visitor, each search, each click to The Underdogs put more and more money in Ultsearch's pockets. With just this site alone, Ultsearch made in the vicinity of a few thousand dollars a day at the hands of unwitting visitors who clicked sponsored links for search terms within his directory. Fortunately, Sarinee was able to buy back his site from Ultsearch for the modest fee of $400-modest because Ultsearch very rarely returns an expired domain to its previous owner.


text:
Q: Is it possible to send a Users Location from an iOS App to a Webserver? I´m new to coding and programming and I have basically no knowledge, but my task is to create an iOS App.
I am working in an app project and the goal is to create an Emergency-App. The idea is that if you open the app, there is one button that you have to press for 2 seconds to send an Emergency call to a Web-Server, including the CoreLocation of the user and if possible also the floor where the user´s currently at. (As mentioned, I have no idea whether it is possible, thats why I am asking)
I also read that some people sent the location with an sms to a phone number.
So if any of you guys could help me on how to send the location to a web server that would be very helpfull! Or if you have an advice for me thats helpfull as well !
Thanks in common!
I haven´t really tried anything yet because i don´t know how.

A: From beginning, First set location Autorization status:
For that you need to specify permission in info.plist.
After the user will allow location services you will be able to get the real location coordinate on real device, if you want to simulate the location you can do it on simulator.
CoreLocation is the framework which Apple provide to access the location service. You sometime need to turn on the location capabilities.
You will have LocationManager class which does all you need. It also has delegate method that will give you all update of location, you can save current location in UserDefault everytime update comes.
When user press button you can send those coordinates from UserDefault


The case of Spiersbridge Property Developments Ltd vMuir Construction Ltd  involved a determination relating to an action alleging breach of a building contract. A bank had paid out an amount demanded by the pursuer under a performance bond and it had to be decided, if the demand on the bond exceeded the sum ultimately due, whether the pursuer was obliged to account for the excess to the bank or to the defender.
The pursuer in this case was a property development company and the defender was a construction company. The parties entered into a building contract in June 2005, in accordance with this contract, the defender was to design and construct a development consisting of warehouses and office space.
This case was centred on the pursuer claiming for alleged delays in completing the works whilst the defender counterclaimed, requesting for an extension of time in which it could complete its obligations.
"…A performance bond in an amount not less than 10% of the Contract Sum in terms the same as the draft performance bond set out in Part Five of this Schedule".
The Bank of Scotland subsequently issued a performance bond. The performance bond was in the form of a letter which was addressed to the pursuers and included substantially the same terms as the draft bond referred to in clause 2.10.2.
Then, in November 2006, the pursuer made a demand requiring the bank to pay £503,193.75 under the bond, which the bank duly paid to the pursuer. The defender stated that it was obliged, under a counter-indemnity it had granted to the bank, to pay that same amount to the bank and that it had duly done so.
"…In the event that… the pursuer should make a call on the bond it would account to the defender for the proceeds of the bond, retaining only the amount equivalent to any loss suffered by the pursuer as a result of the defender's breach of contract, if any".
It was argued that such a term required to be implied as a matter of business efficacy. This meant that the dispute as to whether the defender was in breach of the building contract, as alleged by the pursuer, was yet to be settled.
Despite the fact that a 'proof before answer' had been appointed, the parties also disagreed on whether the pursuer was obliged to account to the defender for that excess, assuming it was found to be entitled to a sum less than it was paid under the bond.
Accordingly, the pursuer argued that its duty to account was owed to the bank and not to the defender. The pursuer's principal concern was that if it made payment of that excess to the defender, it risked being sued for an equal sum by the bank.
§ The banking contract between the defender and the bank pursuant to which the bank agreed to issue the performance bond.
It had to be decided to whom the pursuer should account for the excess and the route to achieve this. It was submitted that the most sensible route was by implication of a term into the bond contract. The term would state that the pursuer would repay the excess to the bank. This would be matched by a corresponding term to be implied into the banking contract under which, if it had already been paid by the defender, the bank would repay the said amount to the defender.
This however raised some potential difficulties. If the term was implied into the building contract where the defender became insolvent, and the pursuer was required to account for the excess to the defender, the payment by the pursuer would go into the pot for the general body of creditors of the defender. This would mean that unless it had already been paid by the defender, the bank would lose out.
Counsel for the defender submitted that the term should be implied in the building contract. If it were the case that it was the bank that could sue on the bond for the excess, the bank would be undertaking the burden of seeking to prove in litigation with the pursuer that the defender was not in breach of contract. Alternatively, that the damage suffered by the pursuer was less than the amount called under the bond.
It was argued that that was not a task a reasonable banker would be particularly willing to undertake, not just because of the difficulty of running such a case, but also due to the fact that it would be expensive.
It would be much better for the bank to be able to rely upon its counter-indemnity from the defender at the time that the bond was called. If the bank had the right of action for recovery of the excess, counsel for the pursuer argued that those difficulties could be overcome by an assignment of the right of action by the bank to the defender. However, this would not work as the terms of the bond prohibited the bank from assigning its rights without the pursuer's consent. Furthermore, if the bank had been paid by the defender pursuant to its counter-indemnity, it would not have suffered any loss and would have no claim to assign.
After much deliberation, the court held that where a demand was made on a performance bond in an amount which was ultimately found to exceed the sum due to the party making the demand, that party was held to be obliged to account for that excess to the opposite contracting party. In the circumstances of this case, the parties had agreed that the obligation on the pursuer to account for any excess must rest upon an implied term in one of the contracts to which it was a party.
This meant that the question then became one of establishing which implication best gave the intended business efficacy to the transaction.
'…In the event that… the pursuer should make a call on the bond it would account to the defender for the proceeds of the bond, retaining only the amount equivalent to any loss suffered by the pursuer as a result of the defender's breach of contract, if any'.
It was held that a term implied into the building contract had none of the disadvantages of involving the bank in the merits of the case. Furthermore, it also allowed for establishing what loss, if any, the pursuer had suffered as a result of the defender's alleged breach of the building contract. This could be determined in litigation or arbitration between the parties to that contract.
The court further held that it was unrealistic to think that the bank would not have agreed with the defender a counter-indemnity in terms of which the defender would in turn indemnify the bank in the like amount upon a call being made upon the bond.
In the event that the call on the bond was excessive, the defender would be out of pocket, not the bank. According to the court, it seemed quite natural that it should be the defender to whom the pursuer had to account for that excess. This did however leave one potential problem. That problem being that if the defender became insolvent after the bond was established by the bank, but before the bank could claim against the defender on the counter-indemnity, then the bank would stand to lose out if it had not taken security.
This potential problem was held to merely be a commercial risk which the bank would decide whether or not to take depending upon its assessment of the defender's creditworthiness. As such, the bank could overcome this problem by refusing to issue the bond, or require some security before agreeing to issue it.
Money becomes an imperative figure when it is about considering any sort of investment or dealing in estate property. Most of the times and for the majority of us, we do not have the true resources and all the necessary information that is required in estate transactions. Therefore, in such situations the best choice is to consult a professional and experienced commercial real estate lawyer who will have all the necessary and precise information during the process.
You may need to approach a lawyer when you are selling or buying commercial property or when you want to make sure that the sale you are getting is fair and legitimately priced. Well, whatsoever is the reason, a lawyer is always the most advisable and sensible option when it comes to dealings in the realty industry. Before we go through the crucial details of commercial real estate lawyers, let us briefly read about real estate law.
This term loosely refers to any piece of land purchased with the full intent of making substantial money on the overall property. Commercial estate can also be used for apartment complexes and industrial production. However, most commonly it is used for restaurants, hotels, shopping malls, office buildings and any other similar property on which the owner of the land expects to mint some money.
What Does a Commercial Real Estate Lawyer Do?
Real estate is basically the sale, purchase, and transfer of homes, buildings or any other commercial property and very strict laws are alleviated for real estate transactions. Thus, a lawyer is typically required for such types of commercial transactions. A lawyer has focused and practiced the real estate law and thus is entitled as a fully licensed attorney. The lawyers handle estate transactions encumbering commercial properties. They also title opinions.
The clients can be property owners, lenders, or developers and can be involved in issues related to property litigation, zoning problems or contract disputes. A lawyer has to render advice to his clients regarding all the paperwork and property transactions. This is one of the most important roles as a commercial lawyer.
Many of the times, established commercial firms hire dedicated lawyers to work for their corporations. Sometimes, these lawyers attain good opportunities and act as general counsel for large scale developers too. However, the job of such attorneys is to sell and buy properties, review the financial documents, file paperwork for corporations, partnerships, documentation for taxes, and communicate with investors regarding the property.
Nevertheless, other attorneys in this domain are also in charge of reviewing the deeds and ensuring the titles are appropriately filed with the right governing policies. They also manage the negotiating leases for various commercial office buildings and shopping centers.
The job of a lawyer is globally recognised and respected as a position of authority and trust. Lawyers' job roles vary significantly depending upon which aspect of law they choose to specialise in. The fact that they generally specialise in only one aspect of law means that you may consult with more than one lawyer in your lifetime depending on the situation; you would need a family lawyer to sort out a divorce whilst a commercial lawyer would ensure that your business is in order.
Family lawyers cover issues such as divorce and child custody. This aspect of law can be very emotive and often refer to sensitive issues therefore lawyers in this field should be approachable and sympathetic. One of the main things to consider when choosing a lawyer in this sector is whether you feel comfortable speaking to them. This is because you will need to share all of the information about the circumstances of the situation which can often be a harrowing experience. Having a lawyer that you are relaxed with should make divulging this information less stressful.
Criminal lawyers work for both the accused party and the victim of crime. When choosing a lawyer if you are accused of a crime ensure that they have a good track record with cases similar to your own, their experience will be invaluable.
If you are a victim of crime, reliving the incident by talking to a lawyer can be a very emotive experience. It is important then that the lawyer gains all of the evidence necessary to build a case against the accused, but also that they are sympathetic and understanding to your feelings and emotions. As with all good family lawyers, criminal lawyers should make you feel comfortable to talk to them about all of the issues surrounding the criminal act.
All businesses, regardless of size, are governed by commercial law. This means that in order to operate, they must be compliant to all aspects of this law. As a result, businesses often use commercial lawyers on a consultancy basis to ensure that their practises are in line with the law. When you employ a commercial lawyer, ensure that you are able to maintain a healthy working relationship with them. This will benefit your company as the more that the lawyer knows about your company, the more they will be able to assist you with decisions and planning for the future. It also means that should you come into difficulty with the law, they will know enough about how your business operates to be able to represent you honestly.
Businesses are responsible for calculating the amount of tax they must pay each year. If this is done incorrectly it can result in businesses overpaying or underpaying tax. Underpayment of tax can result in substantial fines so it is imperative that the tax calculation is accurate and the money reaches the government on time. Tax lawyers can be a useful tool in helping to complete these calculations as they have vast amounts of knowledge about the legal requirements of tax payments. You should choose a tax lawyer that you feel you can trust as they will be dealing with your business finances at least once per year. Also, should you have an issues concerning tax, they will have the inside knowledge to be able to represent you fairly.


This week's metal history lesson celebrates the passionate, dedicated, and massively brave metal bands of Syria. Blackspell, Laxsid, Netherion, Haunted Cellar, Anarchadia, Abidetherin, Theoria, and others present the viewpoints of metalheads raging against total adversity in an active war zone. We salute our brothers in metal halfway across the world and honor the hardship and crisis they have faced for many years now. Know your Roots!


text:
Emmanuel Waine (born 6 October 1996) is a Papua New Guinean professional rugby league footballer who plays as a  and  for the London Broncos in the RFL Championship and Papua New Guinea at international level.

Career
Waine made his international debut for Papua New Guinea in their 24-14 victory over Fiji in the 2022 Pacific Test.

References

External links
PNG Hunters profile

1996 births
Living people
London Broncos players
Papua New Guinea Hunters players
Papua New Guinea national rugby league team players
Papua New Guinean rugby league players
Rugby league forwards

redpajama_set_name: RedPajamaWikipedia


On May 20, 2013 the General Services Administration (GSA) announced the third company to become FedRAMP certified as Amazon Web Services. As part of the program, Amazon has been granted two Agency Authorities to Operate (ATOs) by the U.S. Department of Health and Human Services. CGI Federal and Autonomic Resources are the other two FedRAMP-certified providers.
Run by the GSA, FedRAMP is designed to assist in closing costly federal data centers and outsourcing network operations to shared cloud providers, thereby reducing expenditures and increasing technical flexibility. Once a service goes through the initial FedRAMP authorization process, it will get a stamp of approval that all agencies can use to sign off on the service's ability to meet federal security requirements.
There are three major players in FedRAMP: the cloud service providers (CSPs), the third-party assessment organizations (3PAOs), who will perform an independent audit of the cloud system, and the Joint Authorization Board (JAB), the primary governance and decision-making body for the FedRAMP program. According to the Washington Post, there are about 70 companies in the pipeline for FedRAMP approval.


IITE at Indore, Madurai, Nagpur, Vijayawada, Rajkot, Mangalore & Bhubaneshwar is India's Premier B2C & B2B Travel & Tourism Exhibition. IITE is a One Stop Shop where the upward moving & high earning consumers interact with your Tourism Board, Travel Partners and Affiliated Service providers, thus promoting the beauty and the benefits of the destination. The Event is organized keeping in mind the consumer's Travel & Leisure needs.
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John Robertson spots this week's terrific bargain offered by tool company Screwfix.
Interesting story in The Herald about the young boy on Barra in the Western Isles who flies to Glasgow to train with Greenock Morton. Our correspondent hundreds of miles away on Shetland tells us that staff in the Lerwick office of the BBC were surprised to receive an email from a colleague in Glasgow asking if they could pop across to Barra to cover the story.
We reckon to get from Shetland to Barra would involve an eight-hour journey – by flying via Glasgow.
Says our Shetland man: "It makes a change from some folk in the Beeb who thought that Shetland was in a box floating off Aberdeen."
Our yarn about sports presenter Dougie Donnelly reminds Wilf O'Malley in Inverness of the time Dougie was interviewing Scottish badminton champion Billy Gilliland at the Sports Personality of the Year Awards. Says Wilf: "Dougie with microphone in hand said, 'Well Billy, are you disappointed when you walk down Sauchiehall Street and nobody recognises you?' "Not really' replied Billy. 'I live in London'."
A west end reader tells us her pal was telling friends about how tough she was finding her new fitness regime. She told them: "My personal trainer asked me if I knew that I burned about 70 calories an hour while I was sleeping. I told him, 'Great. Wake me up in 2025'."
We asked for your GP stories, and Hugh Walsh in Dalry tells us: "I wasn't feeling well recently and my wife called the doctor who asked, 'How long have you had these symptoms?' I replied, 'About a week – I didn't want to bother you.' She said, 'People of your generation watched too many John Wayne films'."
The prestigious Wall Street Journal has named Dundee as one of the top ten "Hot destinations" for next year, partly because of the anticipated opening of the V&A Museum of Design. We think though that the new museum may still need a bit of selling locally. Writer Deedee Cuddihy was passing the impressive building on a bus when the boy behind her asked his mum: "What's that museum for?" His mum replied: "I really honestly dinna ken."
Also next year, the National Library of Scotland is marking the centenary of the birth of renowned Scottish writer Muriel Spark. Muriel was still sharp-tongued in her eighties when she was interviewed by The Herald and remarked about fellow authors: "I do know lots of novels that should have been short stories." She also mentioned the snobbery of Brits who had moved out to Tuscany where she then lived. Remarked Muriel: "They buy a tiny cottage with a long avenue of trees leading up to it as if it's Blenheim Palace."
Bringing up teenagers continued. A Pollokshields reader tells us: "My teenage daughter flounced in and announced, 'I hate all my friends!'. So I told her, 'Did we not have a conversation about you being more positive about things?' and she replied, 'Well, I'm positive I hate all my friends'."
Our story about the Brexit pumpkin is challenged by James Miller in Broomhill, Glasgow, who says: "An alternative could be, how to make an EU pumpkin. 1, Britain gives the EU a whole pumpkin. 2, EU says 'Merci' and reciprocates by giving us 3/4 of a pumpkin."


Acute lung injury (ALI) is associated with increased vascular permeability, leukocyte recruitment, and pro‐inflammatory mediator release. We investigated the role of the metalloproteinase ADAM17 in endotoxin‐induced ALI with focus on endothelial ADAM17. In vitro, endotoxin‐mediated induction of endothelial permeability and IL‐8‐induced transmigration of neutrophils through human microvascular endothelial cells required ADAM17 as shown by inhibition with GW280264X or shRNA‐mediated knockdown. In vivo, ALI was induced by intranasal endotoxin‐challenge combined with GW280264X treatment or endothelial adam17‐knockout. Endotoxin‐triggered upregulation of ADAM17 mRNA in the lung was abrogated in knockout mice and associated with reduced ectodomain shedding of the junctional adhesion molecule JAM‐A and the transmembrane chemokine CX3CL1. Induced vascular permeability, oedema formation, release of TNF‐α and IL‐6 and pulmonary leukocyte recruitment were all markedly reduced by GW280264X or endothelial adam17‐knockout. Intranasal application of TNF‐α could not restore leukocyte recruitment and oedema formation in endothelial adam17‐knockout animals. Thus, activation of endothelial ADAM17 promotes acute pulmonary inflammation in response to endotoxin by multiple endothelial shedding events most likely independently of endothelial TNF‐α release leading to enhanced vascular permeability and leukocyte recruitment.
Pulmonary or systemic inflammation can lead to ALI and in severe cases to respiratory distress syndrome and death (25–58%). oedema formation and leukocyte recruitment within the diseased lung tissue are early hallmarks of the inflammatory process. These responses are mediated by the action of cytokines and chemokines, adhesion molecules and junction molecules. Many of these pro‐inflammatory mediator molecules require regulation by limited proteolysis at the cell surface, and in most of the cases, the disintegrins and metalloproteinases ADAM10 and 17 have been made responsible for this shedding process. Therefore, these proteases might represent potential targets for the protein‐ and site‐specific treatment of lung disease. We investigated the influence of ADAM17 on ALI induced by endotoxin exposure with special focus on endothelial ADAM17.
In vitro, endotoxin exposure enhanced expression and activity of ADAM10 and ADAM17, which was associated with increased endothelial permeability and release of the junctional adhesion molecule JAM‐A. Regulation of permeability as well as IL‐8‐induced transmigration of neutrophils through human microvascular endothelial cells required both ADAM10 and 17, as shown by pharmacological inhibition or specific shRNA‐mediated knockdown of the proteases. In vivo, ALI was induced by intranasal endotoxin challenge in combination with metalloproteinase inhibitor treatment or adam17 knockout in endothelial cells. Endotoxin triggered the upregulation of ADAM17 gene expression in the lung, which was abrogated in knockout mice. Induced vascular permeability, oedema formation, release of TNF‐α and IL‐6 and pulmonary leukocyte recruitment were all markedly reduced by inhibitor treatment to block ADAM10 and ADAM17 or endothelial adam17 knockout. This was associated with reduced shedding of JAM‐A and the transmembrane chemokine CX3CL1. Intranasal application of TNF‐α could not restore leukocyte recruitment and oedema formation in knockout animals indicating that these responses are independent of TNF‐α‐release by endothelial cells. Taken together, our in vitro and in vivo experiments show that ADAM17, particular in endothelial cells, acts as a central pro‐inflammatory regulator of pulmonary inflammation. This action appears on different levels, by increase of endothelial permeability involving shedding of junction molecules, enhanced release of inflammatory mediators and facilitation of leukocyte recruitment.
Metalloproteinases of the ADAM family have been recognized as potential therapeutic targets in several diseases. However, due to their broad activity, systemic application of inhibitors may not represent an appropriate treatment strategy. Here, we present first evidence for a critical role of lung endothelial ADAM17 in the development of ALI. Further research is warranted to demonstrate that local ADAM17 inhibition avoiding systemic side effects can represent a promising approach for the treatment of this disease.
Excessive endothelial leakage is a hallmark of acute lung injury (ALI), followed by the accumulation of leukocytes in the alveolar space (Alm et al, 2010; Reiss et al, 2012). Resident and recruited leukocytes fulfil important functions in the clearance of pathogens, but may also damage the endothelial and epithelial cell layer (Gungor et al, 2010; O'Grady et al, 2001; Strieter & Kunkel, 1994). The lung inflammatory response is regulated by a number of soluble and surface‐expressed molecules originating from endothelial cells, smooth muscle cells, epithelial cells and leukocytes (Strieter & Kunkel, 1994; Zhang et al, 2000). Many of these molecules exist as transmembrane molecules that become released from the cell surface by limited proteolysis close to the cell membrane. This process is termed ectodomain shedding and represents a critical regulatory mechanism for the release of cytokines (e.g. TNF‐α), cytokine receptors (e.g. TNFR1 and IL‐6R) (Canault et al, 2006), chemokines (e.g. CXCL16 and CX3CL1; Abel et al, 2004; Hundhausen et al, 2003; Schwarz et al, 2010), adhesion molecules (e.g. l‐selectin) (Walcheck et al, 2006) and surface proteoglycans (e.g. CD44 and syndecans; Haczku et al, 2000; Pruessmeyer et al, 2009). Members of the ADAM (a disintegrin and a metalloproteinase) family, in particular ADAM17, are involved in the ectodomain shedding of these surface molecules (Dreymueller et al, 2011; Pruessmeyer & Ludwig, 2009), and evidence is accumulating that ADAM17 is an important regulator of the acute inflammatory response (Cesaro et al, 2009). In vitro, enhanced ADAM17 expression and activity have been demonstrated in endothelial cells, leukocytes, smooth muscle cells, and epithelial cells upon stimulation with proinflammatory factors such as nitric oxides or cytokines like TNF‐α and IFN‐γ (Bzowska et al, 2004; Canault et al, 2006; Cesaro et al, 2009; Dijkstra et al, 2009). In contrast, in a model of Escherichia coli‐induced peritonitis, mice with a targeted deletion of adam17 in monocytic cells showed improved survival, probably because they were able to recruit more neutrophils to the peritoneal cavity whilst systemic TNF‐α, IL‐1 and IL‐6 levels were reduced (Long et al, 2009).
ADAM17 is widely expressed throughout lung tissue (Dijkstra et al, 2009), and its expression is upregulated in asthma and chronic obstructive pulmonary disease (Dijkstra et al, 2009; Paulissen et al, 2009). The regulation and relevance of lung tissue ADAM17 in ALI is only poorly understood. Additionally, the wide range of functions of ADAM17, evidenced by the fact that ADAM17‐deficient mice die shortly after birth (Peschon et al, 1998), necessitates to define its role in specific cells.
Since endothelial barrier dysfunction is critical for the development of ALI, the present study was designed to investigate the relevance of endothelial ADAM17 activity in acute lung inflammation induced by intranasal instillation of E. coli lipopolysaccharide (LPS). Inhibition and gene silencing experiments with LPS‐stimulated lung microvascular endothelial cells as well as pharmacological inhibition experiments with LPS‐challenged mice suggested a critical role of endothelial ADAM17 for barrier dysfunction and transmigration of leukocytes. This was studied in more detail in mice with targeted deletion of adam17 in endothelial cells. These animals showed diminished cellular infiltration, cytokine secretion and oedema formation upon intranasal LPS‐challenge. Similar observations were made when ALI was induced by TNF‐α, indicating that the beneficial effects of ADAM17 deficiency were not only related to endothelial cell ectodomain shedding of this cytokine, but of other ADAM17 substrates including junction molecules. Our findings show that endothelial ADAM17 activity is critical for the development of ALI.
The role of ectodomain shedding for barrier functions of cultured human microvascular endothelial cells (HMVEC‐L) was analysed by pharmacological inhibition with GW280264X blocking both ADAM10 and ADAM17. The particular contribution of ADAM10 and ADAM17 was addressed by specific lentiviral knockdown of their mRNA expression using shRNA. The knockdown was verified by surface staining of the transmembrane proteases (Fig 1A and B). Treatment of HMVEC‐L with LPS resulted in increased peptide cleavage activity as measured by the proteolysis of a fluorogenic peptide substrate for ADAM10 and ADAM17 (Fig 1C). Addition of the inhibitor GW280264X reduced the cleavage activity by ∼50% in both unstimulated and stimulated HMVEC‐L. The same degree of cleavage inhibition was observed after knockdown of either ADAM10 or ADAM17 (Fig 1C).
Influence of LPS challenge on ADAM10 and ADAM17 activity and mRNA expression in vitro. Data represent means ± SEM (n = 3 per group, C–E). Significance was calculated using one‐way ANOVA followed by the Newman–Keuls post‐test (in D,E) or by two‐way ANOVA followed by the Bonferroni post‐test for double‐treated cells (in C) and is indicated by asterisks (*p < 0.05, **p < 0.01, ***p < 0.001). Asterisks without line indicate significance to the appropriate control.
A‐B. HMVEC‐L were transduced with lentivirus encoding shRNA (LV‐scramble, LV‐antiA10 or LV‐antiA17). Downregulation of ADAM10 (A) or ADAM17 (B) (black line) was analysed by surface staining with antibodies to ADAM10 or ADAM17 compared to isotype controls (light grey tinted) and surface stained scramble transduced cells (black tinted) followed by flow cytometry. Representative histograms of three independent experiments are shown.
C. HMVEC‐L were stimulated for 24 h with LPS (1 µg/ml) or vehicle control (PBS). Cell lysates were analysed for sheddase activity using a fluorogenic peptide cleavage assay. Results were expressed as percentage of sheddase activity in relation to the unstimulated control receiving the vehicle DMSO only (= 100%).
D‐E. HMVEC‐L were stimulated for 0–4 h with LPS (1 µg/ml) or vehicle control (PBS) in the presence of 100 ng/ml LBP and in the absence of serum. ADAM17 (D) and ADAM10 (E) mRNA expression were examined by RT‐qPCR analysis. Data are expressed as change of expression rate compared to control cells.
ADAM10 and ADAM17 can be regulated by altering either enzyme activity or gene transcription. By RT‐qPCR analysis we confirmed that mRNA expression of both proteases is induced already 1 h after stimulation (Fig 1D and E), as also observed for ADAM9 (Supporting Information Fig S1A). After 4 h of stimulation, ADAM17 mRNA expression remained high, whereas the mRNA expression of ADAM10 slightly decreased and that of ADAM9 almost returned to baseline levels. ADAM15 showed delayed induction of mRNA expression after 4 h of LPS challenge (Supporting Information Fig S1B). Analysis of matrix metalloproteinase‐7 (MMP7) mRNA expression revealed an upregulation upon 4 h of stimulation (unpublished observation).
Thus, LPS increases both sheddase activity and mRNA expression of ADAM10 and ADAM17 in microvascular endothelial cells.
Lipopolysaccharide induces paracellular gap formation by shedding of junction proteins, including JAM‐A and cadherins, which are intracellularly linked to β‐catenin (Nonas et al, 2006). Silencing of ADAM17 results in reduced ectodomain shedding of JAM‐A from endothelial and epithelial cells as we have previously described (Koenen et al, 2009). Following LPS treatment (24 h), we observed enhanced permeability measured by TRITC‐dextran diffusion, which was completely suppressed by GW280264X‐treatment (Fig 2A). Silencing of either ADAM10 or ADAM17 suppressed the permeability change to a similar degree as GW280264X (Fig 2A) suggesting that both proteases are required for the control of microvascular permeability under inflammatory conditions. Additionally, immunohistochemistry of β‐catenin as an indicator of established cell junctions showed that LPS treatment (24 h) led to paracellular gap formation, also inhibited by GW280264X‐treatment, without influencing the basal cell morphology (Supporting Information Fig 2).
Role of ADAM10 and ADAM17 in lung microvascular endothelial cells in vitro. HMVEC‐L were transduced with lentivirus encoding shRNA (LV‐scramble, LV‐antiA10 or LV‐antiA17). Data represent means ± SEM (n = 3). Significance was calculated using one‐way ANOVA followed by the Newman‐Keuls post‐test (in B and C) or by two‐way ANOVA followed by the Bonferroni post‐test for double‐treated cells (in A) and is indicated by asterisks (*p < 0.05, **p < 0.01, ***p < 0.001). Asterisks without line indicate significance to the appropriate control.
A. Transduced HMVEC‐L were grown in transwell inserts and stimulated for 24 h with LPS (1 µg/ml) or were left unstimulated (PBS) in the presence or absence of GW280264X (10 µM). Permeability was measured by TRITC‐dextran diffusion. Data are shown as percentage of TRITC‐dextran permeability in relation to the unstimulated DMSO‐treated control (= 100%).
B. HMVEC‐L were stimulated for 4 or 24 h with LPS (1 µg/ml) or were left unstimulated (PBS) in the presence of 100 ng/ml LBP and in the absence of serum. Cells were treated either with GW280264X (10 µM) or with vehicle control (0.1% DMSO). Conditioned media were analysed by immunoblotting followed by densitometric quantification. A representative immunoblot of three independent experiments is shown below the graph.
C. Transduced HMVEC‐L were pre‐treated with or without GW280264X (10 µM) for 1.5 h and examined for IL‐8‐induced (10 ng/ml) transmigration of neutrophils. Experiments were performed with neutrophils from three different donors. Results were expressed as percentage of transmigration in relation to the LV‐scramble‐transduced control (C) receiving no IL‐8.
To investigate whether the LPS‐induced increase in permeability was linked to ADAM10‐ and ADAM17‐mediated shedding of junction molecules, we investigated the release of soluble JAM‐A by LPS‐challenged HMVEC‐L. Release of soluble JAM‐A was 1.3‐fold enhanced by 4 h of LPS stimulation, further increasing to 2.3‐fold by stimulation for 24 h (Fig 2B) and was completely inhibited by GW280264X‐treatment. The knockdown of ADAM10 or ADAM17 by shRNA indicated the involvement of ADAM17 and to a lesser extent of ADAM10 in JAM‐A release (Supporting Information Fig 6).
We further examined whether ADAM10/17 activity might influence transendothelial migration of neutrophils. The inhibitor GW280264X reduced transmigration in response to the neutrophil‐attracting chemokine IL‐8 by 70% (Fig 2C). Silencing of either ADAM10 or ADAM17 alone was sufficient to abrogate transmigration in response to IL‐8 (Fig 2D). Thus, endothelial ADAM10 and ADAM17 are both required for microvascular permeability and for IL‐8‐mediated transmigration of neutrophils in vitro.
Based on the critical role of ADAM10 and ADAM17 for microvascular permeability and transmigration in vitro, we continued to examine their significance in an ALI model in vivo. Intranasal application of LPS strongly increased protein influx into the bronchalveolar lavage (BAL) fluid already 4 h after LPS treatment (Fig 3A, 1.9‐fold), and also increased the tissue wet–dry‐ratio, which was prominent after 24 h (Fig 3B, 1.4‐fold). Both effects, indicating increased vascular permeability and oedema formation, were abrogated by treatment with GW280264X to simultaneously block ADAM10 and 17 (Fig 3A and B).
Effect of ADAM10 and ADAM17 inhibition on permeability, oedema formation and cytokine secretion in LPS‐induced lung inflammation. Mice were intranasally treated with LPS (400 µg/kg) or vehicle control (PBS) with or without the metalloproteinase inhibitor GW280264X (40 µg/kg). Data represent means ± SEM (n = 3 per group). Significance was calculated using one‐way ANOVA followed by the Newman–Keuls post‐test and indicated by asterisks (*p < 0.05, **p < 0.01, ***p < 0.001). Asterisks without line indicate significance to the appropriate control.
A. Whole protein content of BAL fluid was determined 4 h after LPS application.
B. Lung wet–dry‐ratio was determined 24 h after intranasal LPS application.
C‐E. Release of CX3CL1 (C), TNF‐α (D) and IL‐6 (E) into BAL fluid was determined by ELISA 4 h after intranasal application.
We next investigated the release of the chemokine CX3CL1 (fractalkine), which is shed from the cell surface by the activity of ADAM10 and ADAM17 after stimulation with pro‐inflammatory stimuli (Hundhausen et al, 2003). CX3CL1 release was 1.9‐fold increased within the first 4 h of LPS challenge (Fig 3A). This increase was inhibited by intranasal treatment with GW280264X, whereas basal levels in PBS‐challenged controls did not differ.
Release of the pro‐inflammatory cytokine TNF‐α was prominent 4 h after challenge (66‐fold increase, Fig 3D) and was declining after 24 h (unpublished observation). This release was markedly inhibited by intranasal treatment with GW280264X, whereas basal levels in PBS‐challenged controls were not affected. The observation that inhibition of TNF‐α release was not complete may be explained by the fact that TNF‐α is produced at many sites and by many cell types, which may be difficult to access by the inhibitor. Finally, LPS stimulation also induced the release of IL‐6 (28‐fold increase, Fig 3E), which is not an ADAM17 substrate per se. Nevertheless, IL‐6 release was considerably reduced by treatment with the inhibitor, which may reflect the fact that IL‐6 is produced in response to other inflammatory stimuli such as TNF‐α.
These inhibition experiments suggest that either ADAM10 or ADAM17 or both are involved in the regulation of vascular permeability and oedema formation as well as the release of inflammatory mediators in the course of LPS‐induced ALI.
The recruitment of leukocytes to the alveolar space (BAL fluid) and the lung tissue was studied by flow cytometry. LPS challenge increased the total leukocyte number and the number of neutrophils recovered from BAL fluid (Fig 4A and C), which was detectable after 4 h (unpublished observation) and more prominent after 24 h of LPS challenge (11.5‐fold increase of neutrophil BAL count). LPS‐induced neutrophil recruitment was reduced to about 50% by GW280264X‐treatment (Fig 4C). The levels in PBS‐challenged controls remained unaffected by the inhibitor treatment.
Influence of ADAM10 and ADAM17 inhibition on leukocyte recruitment in LPS‐induced lung inflammation. Mice were intranasally treated with LPS or vehicle control (PBS) with or without the metalloproteinase inhibitor GW280264X. After 24 h, lungs were lavaged and the lung tissue was enzymatically disintegrated. Data represent means ± SEM (n = 3 per group). Significance was calculated using one‐way ANOVA followed by the Newman–Keuls post‐test and is indicated by asterisks (*p < 0.05, **p < 0.01, ***p < 0.001). Asterisks without line indicate significance to the appropriate PBS‐treated controls.
A‐D. The total leukocyte cell number (A) as well as the number of macrophages (B) and neutrophils (C) in BAL fluid and the amount of neutrophils in lung tissue suspension (D) were determined by flow cytometry. Results were expressed as cell number per ml BAL fluid (A–C) or as percentage of the total number of lung tissue cells examined (D).
No additional macrophages were recruited upon LPS challenge (24 h). Instead, their recovery from the BAL fluid of LPS‐challenged animals was reduced compared to PBS‐challenged controls (Fig 4B). However, in both PBS‐ and LPS‐challenged animals, the number of macrophages in the BAL fluid was significantly reduced upon application of GW280264X.
For analysis of leukocyte recruitment into interstitial spaces, the lung tissue was disintegrated by enzymatic treatment and analysed for neutrophil content by flow cytometry (see Supporting Information Fig 3 for determination of percentages). Twenty‐four hours after LPS challenge, the number of neutrophils in the lung tissue was 2.2‐fold enhanced (Fig 4D). This response was almost completely suppressed by GW280264X treatment, whereas the levels in PBS‐challenged animals were not significantly affected. These pharmacological studies suggest that either ADAM10 or ADAM17 or both proteases facilitate leukocyte recruitment to the acutely inflamed lung.
We found that LPS challenge increased ADAM17 mRNA in lung tissue (perfused free of blood) of control mice (Fig 5A), whereas ADAM10 expression was not affected. We therefore hypothesized that ADAM17 is more relevant as a regulatory protease in LPS‐induced responses.
Role of endothelial ADAM17 on shedding processes in LPS‐induced lung inflammation. Control and Tie2‐adam17−/− mice were intranasally treated with LPS or vehicle control (PBS). Data represent means ± SEM (n = 3 per group). Significance was calculated using Student's t‐test (in A and B) or using one‐way ANOVA followed by the Newman–Keuls post‐test (in C–F) and is indicated by asterisks (*p < 0.05, **p < 0.01). Asterisks without line indicate significance to the appropriate PBS‐treated controls.
A‐B. After challenge for 24 h, BAL and vascular perfusion, ADAM17 (A) and ADAM10 (B) mRNA expression in lung tissue were examined by RT‐qPCR analysis. Data are expressed as percentage of expression in relation to PBS‐treated controls.
C‐D. Release of soluble JAM‐A into BAL fluid was measured by ELISA 4 and 24 h after intranasal application.
E‐F. Release of soluble CX3CL1 into BAL fluid was determined by ELISA 4 and 24 h after intranasal application.
Based on our in vitro observation with HMVEC‐L that ADAM17 critically controls the induction of endothelial permeability and transmigration, we decided to specifically address the role of endothelial ADAM17 in ALI utilizing adam17−/− mice with Cre recombinase‐mediated deficiency of the protease in endothelial cells (Tie2‐adam17−/−). These mice lack an obvious phenotype, and the absence of ADAM17 mRNA expression in heart and lung endothelial cells was demonstrated (Weskamp et al, 2010). We confirmed the endothelial specificity of ADAM17 knockout in those mice by differential cell sorting of lung tissue and blood cells followed by RT‐qPCR analysis of mRNA expression. Additionally, control mice and Tie2‐adam17−/− mice did not differ with regard to their blood cell composition and the capacity of TNF‐α‐release by alveolar macrophages (for detailed characterization of the mice see Supporting Information Fig 5 and Supporting Information Results).
We then continued to use Tie2‐adam17−/− mice in our model of LPS‐induced lung injury. In these mice, LPS failed to stimulate ADAM17 mRNA expression in lung tissue compared to control animals (Fig 5A), whereas ADAM10 expression did not vary (Fig 5B). Since ADAM17 is involved in the constitutive and induced ectodomain shedding of endothelial JAM‐A, we next determined the level of released soluble JAM‐A in the BAL fluid 4 and 24 h after LPS challenge. In PBS‐treated mice, the release of JAM‐A was generally reduced in Tie2‐adam17−/− mice in comparison to control animals and this difference was more pronounced (up to 30%) when the animals were treated with LPS (Fig 5C and D). In addition, we investigated the level of soluble CX3CL1, which is generated from its transmembrane form via ADAM17‐independent constitutive cleavage and ADAM17‐mediated induced shedding upon cell stimulation (Schwarz et al, 2010). LPS challenge for 4 h increased the release of CX3CL1 into the BAL fluid (Fig 5E, 1.9‐fold) in control animals. As expected, induced CX3CL1 release was abrogated in Tie2‐adam17−/− mice (Fig 5E). CX3CL1 release was generally lower after 24 h of challenge, but was also suppressed in Tie2‐adam17−/− mice (Fig 5F).
Because of the observed deficit of Tie2‐adam17−/− mice in ectodomain shedding of JAM‐A, which is an important component of endothelial tight junctions, we next studied these mice for oedema formation in LPS‐induced ALI. The knockout did not alter the basal level of BAL fluid protein content, but abrogated protein influx in response to LPS (Fig 6A). Similar to the experiments with GW280264X (Fig 3), LPS‐induced oedema formation was completely inhibited in Tie2‐adam17−/− mice (Fig 6B). This was confirmed by histological analysis of the lung tissue. In control mice, LPS treatment increased the thickness of interalveolar septa, whereas Tie2‐adam17−/− mice were protected (Fig 6C and D). Additionally, only LPS‐challenged control mice showed strong tissue damage with loss of parenchymal architecture, whereas the lung tissue of Tie2‐adam17−/− mice stayed intact (Fig 6C).
Role of endothelial ADAM17 for permeability changes and tissue damage in LPS‐induced lung inflammation. Control and Tie2‐adam17−/− mice were intranasally treated with LPS or vehicle control (PBS). Data represent means ± SEM (n = 3 per group for A and B and n = 10 per group for D). Significance was calculated using one‐way ANOVA followed by the Newman–Keuls post‐test and is indicated by asterisks (**p < 0.01, ***p < 0.001). Asterisks without line indicate significance to the appropriate PBS‐treated controls.
A. Whole protein content of BAL fluid was determined 4 h after application.
B. Lung wet–dry‐ratio was examined 24 h after application.
C‐D. After 24 h, 3 µm‐sections of formalin‐fixed and paraffin‐embedded lung tissue were stained with hematoxylin–eosin. Representative images are shown in panel (C). Ten images per animal were analysed for the thickness of interalveolar septa using AixoVision software (D).
Cytokine determination within BAL fluid revealed that LPS‐challenged Tie2‐adam17−/− mice showed reduced release of IL‐6 (Fig 7A, 3.9‐fold) and TNF‐α (Fig 7B, 1.6‐fold), whereas no effect was observed in PBS‐challenged mice. Interestingly, the 37‐fold increased release of the neutrophil attracting chemokine KC (Fig 7C, 36.5‐fold) in LPS‐challenged mice did not differ between the two groups.
Role of endothelial ADAM17 for cytokine secretion in LPS‐induced lung inflammation. Control and Tie2‐adam17−/− mice were intranasally treated with LPS or vehicle control (PBS). Data represent means ± SEM (n = 3 per group). Significance was calculated using one‐way ANOVA followed by the Newman–Keuls post‐test and is indicated by asterisks (*p < 0.05, **p < 0.01, ***p < 0.001). Asterisks without line indicate significance to the appropriate PBS‐treated controls.
A‐C. After 4 h of challenge, release of IL‐6 (A), TNF‐α (B) and KC (C) into BAL fluid was determined by ELISA measurement.
D. Control and Tie2‐adam17−/− mice were compared for TNF‐α‐induced lung inflammation. The wet‐dry‐ratio of lung tissue was determined after 24 h of intranasal TNF‐α‐challenge.
Soluble TNF‐α is generated by ADAM17 and this cytokine is a mediator of ALI (Mazzon & Cuzzocrea, 2007; Smith et al, 1998; Song et al, 2001). Since soluble TNF‐α levels were only moderately reduced in the Tie2‐adam17−/− mice, we questioned whether the protection of vascular leakage and tissue damage, which was seen in the endothelial adam17‐deficient mice, could be explained by the reduced release of TNF‐α. Control and Tie2‐adam17−/− mice were studied for oedema formation after intranasal TNF‐α challenge. The TNF‐α‐induced increase in lung tissue wet–dry‐ratio was almost completely abrogated (Fig 7D) in Tie2‐adam17−/− mice. Therefore, even in the presence of surplus soluble TNF‐α, mice lacking endothelial ADAM17 are still protected from ALI.
Given the fact that LPS‐induced TNF‐α release was only moderately reduced and production and release of KC was not affected in mice with endothelial ADAM17 deficiency, we asked whether the almost completely restored vascular permeability could still be associated with altered recruitment of leukocytes in the lungs of these mice. Both the total leukocyte and neutrophil number in BAL fluid 24 h after LPS challenge were significantly reduced in Tie2‐adam17−/− mice (Fig 8A and B), whereas the cell number in BAL fluid upon PBS challenge did not differ. Neutrophils in the lung tissue of LPS‐treated mice were strongly reduced in Tie2‐adam17−/− mice (Fig 8D), similar to mice treated with GW280264X (Fig 4). This was confirmed by neutrophil counts in histological sections (Fig 8E).
Role of endothelial ADAM17 for leukocyte recruitment in LPS‐induced lung inflammation. Control and Tie2‐adam17−/− mice were intranasally treated with LPS or vehicle control (PBS). Data represent means ± SEM (n = 3 per group for A–D and F and n = 10 per group for E). Significance was calculated using one‐way ANOVA followed by the Newman–Keuls post‐test and is indicated by asterisks (*p < 0.05, **p < 0.01, ***p < 0.001). Asterisks without line indicate significance to the appropriate PBS‐treated controls.
A‐D. Twenty‐four hours after application, the number of total leukocytes (A), neutrophils (B) and macrophages (C) in BAL fluid and the amount of neutrophils in lung tissue suspension (D) were determined by flow cytometry. Results were expressed as cell number per ml BAL fluid (A–C) or as percentage of the total number of lung tissue cells investigated (D).
E. Ten images of hematoxylin–eosin stained 3 µm‐sections per animal were analysed for invading neutrophils using the AixoVision software.
F. Control and Tie2‐adam17−/− mice were compared for TNF‐α‐induced lung inflammation. The amount of neutrophils in BAL fluid was determined by flow cytometry after 24 h of intranasal TNF‐α‐challenge.
Since Tie2‐adam17−/− animals showed reduced ectodomain shedding of the transmembrane chemokine CX3CL1, which mediates transendothelial recruitment of monocytic cells, we further investigated the number of macrophages in the BAL fluid 24 h after LPS challenge. Remarkably, in both PBS‐ and LPS‐treated groups, the number of macrophages was reduced in Tie2‐adam17−/− animals (Fig 8C), suggesting that endothelial ADAM17 may regulate the number of macrophages in the alveolar space.
Notably, also in TNF‐α‐treated Tie2‐adam17−/− mice, neutrophil recruitment to BAL fluid was reduced up to 50% (Fig 8F) in comparison to TNF‐α‐treated control mice. Therefore, LPS‐induced lung injury does not appear to particularly depend on endothelial TNF‐α release by ADAM17. Instead, endothelial ADAM17 regulates the inflammatory response at multiple levels, including the shedding of junctional adhesion molecules, and thereby affecting endothelial permeability and the generation and release of various inflammatory cytokines and chemokines such as TNF‐α, IL‐6 and CX3CL1 that promote leukocyte recruitment.
Endothelial barrier dysfunction is a hallmark of ALI. Here, we provide in vitro and in vivo evidence that endothelial ADAM17 plays a critical role in this process. To this end, we used human microvascular endothelial cells of the lung (HMVEC‐L) in culture and endothelium‐specific ADAM17 knockout mice (Tie2‐adam17−/−) to demonstrate that endothelial ADAM17 is critically involved in endothelial barrier dysfunction, oedema formation, cytokine release as well as leukocyte transmigration and recruitment in response to LPS. As shown by successful treatment with GW280264X, ADAM17 may be a worthwhile therapeutic target for the treatment of ALI.
Lipopolysaccharide treatment in vitro and in vivo increased the ADAM17 mRNA expression in cultured lung endothelial cells and in the lungs of mice, respectively. The upregulation was associated with an increase in ADAM17 activity and cleavage of ADAM17 substrates within a similar time frame. Dysregulation of ADAM17 on the level of gene transcription has been reported for other inflammatory diseases including atherosclerosis (Canault et al, 2006). In fact, the protease was found to play a critical pathogenic role in various models of inflammation (Canault et al, 2010; Chalaris et al, 2010; Horiuchi et al, 2007; Li et al, 2006; Long et al, 2009). Moreover, ALI was reduced by metalloproteinase inhibitors (PKF242‐484, PKF241‐466, Y‐41654; Shimizu et al, 2009; Trifilieff et al, 2002). These inhibitors could not distinguish between ADAM17 and matrix metalloproteinases or other ADAM proteases, and did not include early and late time point measurements. Therefore, these pharmacological studies provided no conclusive evidence for a critical role of ADAM17 in ALI and could not address a cell‐type specific contribution of the protease. Genetic studies in this area are complicated by the fact that adam17 total knockout leads to rapid postnatal lethality (Horiuchi et al, 2007; Peschon et al, 1998). As vascular permeability caused by endothelial barrier changes is central for the development of ALI, we used Tie2‐adam17−/− mice that lack ADAM17 in their endothelial cells only (Weskamp et al, 2010) to investigate the contribution of ADAM17 in more detail.
The Tie2‐adam17−/− mice have been characterized thoroughly for the lack of adam17 in endothelial cells (Weskamp et al, 2010). It has been argued that the Tie2‐promoter might be also active in monocytes and neutrophils (Pucci et al, 2009) but we ruled this possibility out by showing normal ADAM17 mRNA expression in peripheral blood cells of Tie2‐adam17−/− mice as well as normal TNF‐α production in alveolar macrophages isolated from Tie2‐adam17−/− mice. Taken together, we conclude that Tie2‐adam17−/− mice lack ADAM17 in endothelial cells only.
Acute lung injury was studied in a model of LPS‐induced pulmonary inflammation that resembles the reaction of the human lung to endotoxin exposure (Matute‐Bello et al, 2008; Nonas et al, 2006). In this model, animals treated with the dual ADAM10/17 inhibitor GW280264X, but also Tie2‐adam17−/− mice were protected on many levels: cytokine release, permeability, oedema formation and neutrophil recruitment. These results indicate a critical role of endothelial ADAM17 in the development of ALI.
There are several ways how ADAM17 substrates could contribute to barrier dysfunction, cytokine release and cell recruitment in ALI. Increased vascular permeability is caused by paracellular gaps that are formed by subsequent reorganization of the cytoskeleton and redistribution of junctional proteins including JAM‐A, which is involved in the regulation of brain endothelial and intestinal epithelial permeability (Haarmann et al, 2010; Laukoetter et al, 2007). ADAM17 also regulates leukocyte rolling along activated endothelium by shedding l‐selectin (Ponnuchamy & Khalil, 2008) and leukocyte transmigration by cleaving CX3CL1 (Schwarz et al, 2010), ICAM‐1 and VCAM‐1 (Pruessmeyer & Ludwig, 2009; Singh et al, 2005) as well as JAM‐A (Koenen et al, 2009). Inhibition of these shedding events results in reduced or abrogated leukocyte transmigration. Indeed, we observed reduced release of soluble JAM‐A from LPS‐stimulated HMVEC‐L upon GW280264X treatment or shRNA‐mediated knockdown of ADAM17 as well as reduced release of JAM‐A and CX3CL1 in Tie2‐adam17−/− mice, exemplifying the importance of ADAM17 for endothelial barrier dysfunction and leukocyte recruitment. Thus, multiple cleavage events within the lung and here particularly in endothelial cells contribute to the regulation of vascular permeability and neutrophil transmigration in vitro and in vivo, and therefore no single ADAM17 substrate can be held responsible for this process. Moreover, the pro‐migratory function of endothelial ADAM17 might not be limited to neutrophils, but might also account for macrophages even in the absence of inflammation as indicated by the finding that the basal number of macrophages in the lungs was reduced in Tie2‐adam17−/− mice.
While we believe that the regulation of endothelial barrier function accounts for the majority of the protection by ADAM17 inhibition against LPS‐induced lung injury, we also observed reduced levels of TNF‐α and IL‐6 in both mice with pharmacological inhibition of ADAM10/17 or genetic ablation of adam17 in endothelial cells. Among these, in particular TNF‐α, a well‐known substrate of ADAM17, is known for its role in endotoxin‐induced lung injury and other forms of ALI (Mazzon & Cuzzocrea, 2007; Schnyder‐Candrian et al, 2005; Smith et al, 1998; Song et al, 2001). However, in our study, the reduction in TNF‐α levels by pharmacological inhibition of ADAM10/17 or genetic ablation of adam17 in endothelial cells was less pronounced compared to studies using rather unspecific inhibitors, where the LPS‐induced TNF‐α levels were almost abolished (Trifilieff et al, 2002). This can be explained by the fact that TNF‐α is predominantly derived from epithelial cells and alveolar macrophages rather than endothelial cells (Ward, 2003) and that other proteases potentially contribute to TNF‐α cleavage (Armstrong et al, 2006). ADAM9, ADAM10 and ADAM19 are capable of cleaving TNF in vitro and also MMP7 as well as proteinase‐3 can shed TNF in vivo. Besides, recruited neutrophils further secrete proinflammatory cytokines and tissue damaging substances (Yoshikawa et al, 2004). Therefore, reduced leukocyte recruitment in mice with pharmacological inhibition of ADAM10/17 or genetic ablation of adam17 in endothelial cells is a likely explanation for the observed decrease in cytokine production such as TNF‐α and IL‐6 (Ward, 2003). Notably, we observed that endothelial hyperpermeability and leukocyte recruitment were reduced in Tie2‐adam17−/− mice regardless of whether ALI was induced by LPS or TNF‐α, indicating that this protection seen in Tie2‐adam17−/− mice is independent of TNF‐α‐release by endothelial ADAM17.
Taken together, ADAM17 mediates a number of shedding events that influence several components of acute lung inflammation including vascular leakage, leukocyte recruitment and cytokine release. Particularly in endothelial cells, ADAM17 appears to act as a central regulator in pulmonary inflammation. We conclude that inhibition of ADAM17, possibly locally to reduce systemic side affects, may be a promising approach for the treatment of ALI.
Antibodies for flow cytometry and immunohistochemistry as well as cytokines and commercially available inhibitors are described in the supplement. The metalloproteinase inhibitor GW280264X was synthesized and assayed for inhibition of human and mouse ADAM17 and ADAM10 as described (Ludwig et al, 2005; Rabinowitz et al, 2001).
Culture of HMVEC‐L (Lonza, Belgium) and alveolar macrophages are described in the supplements. Vectors for lentiviral knockdown of adam10 and 17 have been previously described (Schwarz et al, 2010). Lentiviral transduction and immunohistological examination of HMVEC‐L for β‐catenin are detailed in the supplements.
The mRNA levels were quantified by RT‐qPCR analysis. RNA was extracted using RNeasy Kit (Qiagen, Hilden, Germany), reverse transcribed using RevertAid First Strand cDNA Synthesis Kit (Fermentas, St. Leon‐Rot, Germany) and PCR reactions performed using LightCycler®480 SYBR Green I Master (Roche, Munich Germany) according to the manufacturer's protocols. Primers and amplification protocols are included in supplements.
ADAM activity in cell lysates was measured as cleavage of a fluorogenic peptide‐based substrate mimicking the α‐cleavage site of amyloid‐precursor protein. Changes in ADAM10/17 activity were also measured in terms of JAM‐A release. For details see Supporting Information.
Neutrophils were isolated and analysed for transmigration through HMVEC‐L as described (Schwarz et al, 2010). Transmigrated cells were quantified by measurement of endogenous glucuronidase activity as detailed in Supporting Information.
Permeability was determined by TRITC‐dextran diffusion as described before and within Supporting Information (Koenen et al, 2009).
Animal experiments were approved by the local authorities and performed with 6–8 week old Tie2‐adam17−/− mice or wild type C57BL/6 mice (Janvier, Le Genest sur l'Isle, France), respectively. Tie2‐adam17−/− mice for targeted disruption of adam17 in endothelial cells were generated and characterized in the laboratory of Carl Blobel (Cornell University, New York, USA) and were kindly provided for this study (Weskamp et al, 2010).
Mice were anesthesized by intraperitoneal injection of ketamin (100 mg/kg) and xylazin (5 mg/kg). LPS (400 µg/kg in PBS) or PBS as control were intranasally instilled in the presence or absence of GW280264X (40 µg/kg, dissolved in DMSO, end concentration 0.06%) or vehicle control (0.06% DMSO). After 4 or 24 h, BAL fluid was collected for determination of protein influx, ELISA measurements of cytokine release and flow cytometric analysis of cell recruitment. Lung tissue was used for determination of wet–dry‐ratio and disintegrated for analysis of leukocyte infiltration. TNF‐α (200 µg/kg in PBS) was intranasally instilled in the presence of 0.06% DMSO, and lung inflammation examined after 24 h. For detailed description see Supporting Information.
Quantitative data are shown as mean ± SEM from three independent experiments/cell isolates/animals unless indicated otherwise. Statistical analysis was performed using GraphPad PRISM 5.0 (GraphPad Software, La Jollla, CA). For details see figure legends and the Supporting Information.
DD, CM, SU and AL designed study and wrote manuscript; DD and AL analysed data; DD, TK, JP and FMH performed experiments; KH provided vital technique and intellectual input.
We thank Carl Blobel for providing Tie2‐adam17−/− mice, generous support, and constructive discussions. We thank Felix Heymann and Frank Tacke (Medizinische Klinik III, RWTH Aachen University Hospital) for cell sorting. We thank Nadine Ruske and Melanie Esser for expert technical assistance, and Julian Schumacher for lentivirus production. The work has been supported by the IZKF Aachen of the RWTH Aachen University and the German Research Foundation Lu869/5‐1.


*DISCLOSURE* Thank you to Sleep Zzz Pillow for providing product samples in order to facilitate this post. All thoughts are strictly my own.
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When it comes to pet products, there aren't regulations like there are for human products. Unfortunately, without regulations, the only information we see when purchasing pet products is marketing.
I am not sure if it's a blessing or a curse, but, I instinctively always want to dig deeper. It is definitely a blessing for those of you who have the Sydney Sleep Mattress. For me, however, my brain won't stop researching the next pet product that must be replaced.
How often do most people really do the research before making purchases for their furry family members? I don't blame anyone for reading "natural" on a package and wanting to run with it. When you dig a little deeper, it might end up shaking up your world. None of us have time for that.
I am on a mission to introduce the Sydney Sleep Mattress worldwide. I have done the homework so you don't have to. You can rest assured that I will be the one losing sleep over your pet products so you don't have to.
I talk about the hemp aspect of the Sydney Sleep Mattress so often, it's hard to believe, but, that is not what makes this bed the best. It's the combination of what went into the design and the materials. You have to have both.
I am not the first person to create a dog bed made of hemp. I am the first that didn't stop there. There isn't a bed on the market that can be compared to the Sydney Sleep Mattress apples to apples.
The first 8 beds that came up are not made of hemp at all. In fact, I know someone who purchased the $400 bed that popped up first and she swore to me it was hemp. This was several months ago. I did the research. This bed is "proprietary waterproof fabric" with a foam inner. It is not hemp and more than likely whatever makes it waterproof also makes it off-gas. She also purchased a Sydney Sleep Mattress. Guess which bed the dog chose?
After scrolling through all the non-hemp beds that were mostly made of materials that can possibly off-gas and cause cancer and other diseases, I found some beds actually made of hemp. They are still the same beds that were out there years ago when I was researching for beds myself.
The first one is a very well known pet product company in the boutique and alternative pet product stores. Most of their toys are made of recycled materials which is what put them on the map. They offered a bed made of hemp and you can still find it when you Google, but, they don't actually offer it anymore. Hemp is very expensive. Which means profit margins are tough to hit for retail stores. Also, this bed was really just a glorified pillow. It was not supportive and didn't have much else to offer. To me, that is a waste of expensive fabric.
Then, there is a company making multiple hemp products for dogs. I love that their bed's core is also made of natural fibers, however, it is just a pillow. It is lacking in support. It is expensive and it is not going to be nearly as durable as the Sydney Sleep Mattress. The price tag is over $80 for a small bed that is really just a pillow. The large is $140 and still smaller than the Sydney Sleep Mattress.
I didn't post this to rag on other products or to complain about the pet product industry. I think it's important that consumers know what to look for. I am particularly passionate about dog beds because the industry is so flooded with garbage and dogs spend the majority of their lives sleeping.
If there is an area as important as your dog's nutrition, it is their sleep environment. Invest in their health and well-being. It's something you will never regret.
The introductory price is still good. Use code INTRO and save $130.


text:
Q: When should I specify REML = FALSE in lmer()? I am running a basic Mixed Effect Model with lmer(), in R.
Let say I have 2 within-subject conditions. In each condition the subject provides one measure.
lmer(measure ~ condition + (1|subject),
      REML = TRUE, data = My_data)

The argument REML is TRUE by default. Yet, in several examples I read, people set it as FALSE.
According to the documentations "logical scalar - Should the estimates be chosen to optimize the REML criterion (as opposed to the log-likelihood)?"
Are there real differences in the estimates of fixed or random effects when I use one method or another. When should REML be TRUE? When should it be FALSE?

A: Based on the comment above, a good answer can be found here:
https://stats.stackexchange.com/questions/272633/how-to-decide-whether-to-set-reml-to-true-or-false/272654#272654
To cite the author:
"It's generally good to use REML, if it is available, when you are interested in the magnitude of the random effects variances, but never when you are comparing models with different fixed effects via hypothesis tests or information-theoretic criteria such as AIC."


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When the battery is fully charged, the indicator of the charger turns green.
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12 months Manufacturer's Warranty . Buy with confidence!
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2. If there is any damage, please don't dismantle it. Sent it back to us for repair, and we will give you response or send off within one week after receiving.


text:
Cosmos » Animals » Foxy researchers identify gene for tameness
6 August 2018 /
Andrew Masterson
Foxy researchers identify gene for tameness
Experiment started more than 50 years ago starts to produce results.
Over many generations, foxes can be bred so that hanging out with humans and being generally cute is their dominant characteristic. Credit: Jane Khomi/ Getty Images
The world's oldest, and perhaps only, colony of friendly foxes may have revealed the gene driving tameness – a discovery with potential implications for research into human neurological disorders.
Keeping red foxes (Vulpes vulpes) in captivity on a large scale began in Canada in the late nineteenth century as farmers realised there was money to be made in the animals' pelts. However, the foxes themselves weren't domesticated, and always reacted to their human captors with fear or aggression (perhaps with good reason).
In 1959, however, researchers motivated more by curiosity than cash started a unique fox farm at the Institute of Cytology and Genetics of the Russian Academy of Sciences in St Petersburg. The scientists were interested in the mechanics of domestication and consistently selected foxes that showed affection for humans, cross-breeding them over more than 50 generations.
The result is a still-thriving, and still pure, population of extremely friendly animals.
In the 1960s, researchers at the institute set out to see if they could create the opposite – a population of foxes that reacted with unwavering aggression towards humans. They succeeded, again using conventional cross-breeding methods, and that population has now been around for more than 40 generations.
And as befits decent science, of course, the institute also hosts a third, control fox population that has not been bred to highlight any particular behavioural trait.
Given the integrity of the three populations, a team of researchers led by Anna Kukekova from the University of Illinois at Urbana-Champaign in the US reasoned that differences in the genomes of the three populations should indicate genes that are in some way involved in the expression of tameness or aggression.
Sequencing genomes from members of the nice, nasty and control populations, the scientists found 103 regions that differed between the three groups – and were therefore the result of selection pressures. Some of these included genes that are associated with neurological disorders in humans, including autism and bipolar, while others are known to code for tame behaviours in dogs.
However, they also identified what they termed "a strong positional candidate gene for tame behaviour". This was a gene called SorCS1, which is involved in coding for proteins that regulate communication between neurons. In humans it is also associated with narcolepsy.
The St Petersburg fox colonies were established more than half a century ago, and given the turbulent history of Russia it perhaps seems remarkable that they have survived and been cared for in the interim.
The current study – published in the journal Nature Ecology & Evolution – was funded by research organisations in the US, Russia, China and Germany. With that amount of interest worldwide, it seems that the survival of the animals is pretty much guaranteed for the next half century.
"Decades of documented selection that have resulted in dramatic differences in the behaviour of tame and aggressive foxes render these populations valuable to genomic studies of behaviour," the researchers conclude.
Originally published by Cosmos as Foxy researchers identify gene for tameness
Andrew Masterson is a former editor of Cosmos.
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Q: Talend - Send HTML table - Guarantee that we haven't Wrap Text I'm using Talend Big Data Platform to send a CSV as HTML table by email. To build the HTML code I've a JavaRow with this:
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With this code my table is wrapping my columns and I get my columns names and the values cut in half.
What I want is to wrap text in order to fit the column width and to not have any cut off in cells.
What I am doing wrong?
Many thanks!


12.28.2010 Story by Jason Billington KARBALA PROVINCE, Iraq – As Shiite pilgrims slowly filtered out of religious shrines in Karbala Dec. 17, the quiet passing of the Muslim religious holiday of Ashoura made a loud statement about the capabilities of Iraqi security forces in the area.
As many as 4 million pilgrims took part in the 2010 Ashoura, including 200,000 foreigners from 24 separate countries, according to Iraqi estimates. The security of Ashoura participants and the areas surrounding holy shrines in Karbala was the sole responsibility of Iraqi security forces, who assumed the security lead for the country as part of Operation New Dawn Sept. 1.
"the tenth day," commemorates the death of Imam Al-Hussein, the grandson of the Islamic Prophet Muhammad at the Battle of Karbala in the seventh century. Ashoura is particularly significant to Shia Muslims, who make pilgrimages in order to congregate at holy shrines in Muslim holy cities like Karbala and Najaf. The influx of vast numbers of pilgrims into Iraq presents unique security challenges to the Iraqi security forces, specifically for the crowds of people who gather around the Imam Al-Hussein and Imam Al-Abbas shrines at Karbala.
In previous years the holiday has occasionally been marred by insurgent attempts to disrupt Ashoura gatherings and destabilize the country. In March 2004, two bombs in Baghdad and Karbala killed more than 170 people. Clashes between sectarian groups in January 2008 during Ashoura resulted in the deaths of 263 people in Karbala. No such violence could be seen on the streets of Karbala this year.
U.S. forces assumed a supporting role in the security for Ashoura, advising key ISF leaders and providing humanitarian assistance for participants during the commemorations.
While Ashoura was visibly marked by throngs of people honoring Muslim religious traditions in peace, the Iraqi military and police were busy behind the scenes responding to insurgent threats.
"Ashoura was a very successful operation," said Lt. Gen. Othman Al Ghanimi, commander of the 8th Iraqi Army and charged as the lead in Karbala during the security operations surrounding the holiday. Eighty suspected insurgents were captured and numerous explosives were dismantled by members of Iraqi security forces in and around Karbala, said Othman.
The strength of Iraqi security forces has become paramount as Jan. 1 marks the last year of U.S. forces involvement under the existing security agreement in Iraq.
As a significant Shiite Muslim religious holiday ended Dec. 18, it was not the tested crisis response of Iraqi security forces that marked its successful passing, but the lack of any crisis at all.