text:
using Avalonia.Controls.Metadata;
using Avalonia.Controls.Primitives;
using Avalonia.Controls.Shapes;
using Avalonia.Controls.Templates;
using Avalonia.Data;
using Avalonia.Layout;
using System;
using System.Globalization;

namespace Avalonia.Controls
{
    /// <summary>
    /// A control to allow the user to select a time.
    /// </summary>
    [TemplatePart("FirstColumnDivider",      typeof(Rectangle))]
    [TemplatePart("FirstPickerHost",         typeof(Border))]
    [TemplatePart("FlyoutButton",            typeof(Button))]
    [TemplatePart("FlyoutButtonContentGrid", typeof(Grid))]
    [TemplatePart("HourTextBlock",           typeof(TextBlock))]
    [TemplatePart("MinuteTextBlock",         typeof(TextBlock))]
    [TemplatePart("PeriodTextBlock",         typeof(TextBlock))]
    [TemplatePart("PickerPresenter",         typeof(TimePickerPresenter))]
    [TemplatePart("Popup",                   typeof(Popup))]
    [TemplatePart("SecondColumnDivider",     typeof(Rectangle))]
    [TemplatePart("SecondPickerHost",        typeof(Border))]
    [TemplatePart("ThirdPickerHost",         typeof(Border))]
    [PseudoClasses(":hasnotime")]
    public class TimePicker : TemplatedControl
    {
        /// <summary>
        /// Defines the <see cref="MinuteIncrement"/> property
        /// </summary>
        public static readonly DirectProperty<TimePicker, int> MinuteIncrementProperty =
            AvaloniaProperty.RegisterDirect<TimePicker, int>(nameof(MinuteIncrement),
                x => x.MinuteIncrement, (x, v) => x.MinuteIncrement = v);

        /// <summary>
        /// Defines the <see cref="Header"/> property
        /// </summary>
        public static readonly StyledProperty<object> HeaderProperty =
            AvaloniaProperty.Register<TimePicker, object>(nameof(Header));

        /// <summary>
        /// Defines the <see cref="HeaderTemplate"/> property
        /// </summary>
        public static readonly StyledProperty<IDataTemplate> HeaderTemplateProperty =
            AvaloniaProperty.Register<TimePicker, IDataTemplate>(nameof(HeaderTemplate));

        /// <summary>
        /// Defines the <see cref="ClockIdentifier"/> property
        /// </summary>
        public static readonly DirectProperty<TimePicker, string> ClockIdentifierProperty =
           AvaloniaProperty.RegisterDirect<TimePicker, string>(nameof(ClockIdentifier),
               x => x.ClockIdentifier, (x, v) => x.ClockIdentifier = v);

        /// <summary>
        /// Defines the <see cref="SelectedTime"/> property
        /// </summary>
        public static readonly DirectProperty<TimePicker, TimeSpan?> SelectedTimeProperty =
            AvaloniaProperty.RegisterDirect<TimePicker, TimeSpan?>(nameof(SelectedTime),
                x => x.SelectedTime, (x, v) => x.SelectedTime = v,
                defaultBindingMode: BindingMode.TwoWay);

        // Template Items
        private TimePickerPresenter? _presenter;
        private Button? _flyoutButton;
        private Border? _firstPickerHost;
        private Border? _secondPickerHost;
        private Border? _thirdPickerHost;
        private TextBlock? _hourText;
        private TextBlock? _minuteText;
        private TextBlock? _periodText;
        private Rectangle? _firstSplitter;
        private Rectangle? _secondSplitter;
        private Grid? _contentGrid;
        private Popup? _popup;

        private TimeSpan? _selectedTime;
        private int _minuteIncrement = 1;
        private string _clockIdentifier = "12HourClock";

        public TimePicker()
        {
            PseudoClasses.Set(":hasnotime", true);

            var timePattern = CultureInfo.CurrentCulture.DateTimeFormat.ShortTimePattern;
            if (timePattern.IndexOf("H") != -1)
                _clockIdentifier = "24HourClock";
        }

        /// <summary>
        /// Gets or sets the minute increment in the picker
        /// </summary>
        public int MinuteIncrement
        {
            get => _minuteIncrement;
            set
            {
                if (value < 1 || value > 59)
                    throw new ArgumentOutOfRangeException("1 >= MinuteIncrement <= 59");
                SetAndRaise(MinuteIncrementProperty, ref _minuteIncrement, value);
                SetSelectedTimeText();
            }
        }

        /// <summary>
        /// Gets or sets the header
        /// </summary>
        public object Header
        {
            get => GetValue(HeaderProperty);
            set => SetValue(HeaderProperty, value);
        }

        /// <summary>
        /// Gets or sets the header template
        /// </summary>
        public IDataTemplate HeaderTemplate
        {
            get => GetValue(HeaderTemplateProperty);
            set => SetValue(HeaderTemplateProperty, value);
        }

        /// <summary>
        /// Gets or sets the clock identifier, either 12HourClock or 24HourClock
        /// </summary>
        public string ClockIdentifier
        {
            get => _clockIdentifier;
            set
            {
                if (!(string.IsNullOrEmpty(value) || value == "" || value == "12HourClock" || value == "24HourClock"))
                    throw new ArgumentException("Invalid ClockIdentifier");
                SetAndRaise(ClockIdentifierProperty, ref _clockIdentifier, value);
                SetGrid();
                SetSelectedTimeText();
            }
        }

        /// <summary>
        /// Gets or sets the selected time. Can be null.
        /// </summary>
        public TimeSpan? SelectedTime
        {
            get => _selectedTime;
            set
            {
                var old = _selectedTime;
                SetAndRaise(SelectedTimeProperty, ref _selectedTime, value);
                OnSelectedTimeChanged(old, value);
                SetSelectedTimeText();
            }
        }

        /// <summary>
        /// Raised when the <see cref="SelectedTime"/> property changes
        /// </summary>
        public event EventHandler<TimePickerSelectedValueChangedEventArgs>? SelectedTimeChanged;

        protected override void OnApplyTemplate(TemplateAppliedEventArgs e)
        {
            if (_flyoutButton != null)
                _flyoutButton.Click -= OnFlyoutButtonClicked;

            if (_presenter != null)
            {
                _presenter.Confirmed -= OnConfirmed;
                _presenter.Dismissed -= OnDismissPicker;
            }
            base.OnApplyTemplate(e);

            _flyoutButton = e.NameScope.Find<Button>("FlyoutButton");

            _firstPickerHost = e.NameScope.Find<Border>("FirstPickerHost");
            _secondPickerHost = e.NameScope.Find<Border>("SecondPickerHost");
            _thirdPickerHost = e.NameScope.Find<Border>("ThirdPickerHost");

            _hourText = e.NameScope.Find<TextBlock>("HourTextBlock");
            _minuteText = e.NameScope.Find<TextBlock>("MinuteTextBlock");
            _periodText = e.NameScope.Find<TextBlock>("PeriodTextBlock");

            _firstSplitter = e.NameScope.Find<Rectangle>("FirstColumnDivider");
            _secondSplitter = e.NameScope.Find<Rectangle>("SecondColumnDivider");

            _contentGrid = e.NameScope.Find<Grid>("FlyoutButtonContentGrid");

            _popup = e.NameScope.Find<Popup>("Popup");
            _presenter = e.NameScope.Find<TimePickerPresenter>("PickerPresenter");

            if (_flyoutButton != null)
                _flyoutButton.Click += OnFlyoutButtonClicked;

            SetGrid();
            SetSelectedTimeText();

            if (_presenter != null)
            {
                _presenter.Confirmed += OnConfirmed;
                _presenter.Dismissed += OnDismissPicker;

                _presenter[!TimePickerPresenter.MinuteIncrementProperty] = this[!MinuteIncrementProperty];
                _presenter[!TimePickerPresenter.ClockIdentifierProperty] = this[!ClockIdentifierProperty];
            }
        }

        private void SetGrid()
        {
            if (_contentGrid == null)
                return;

            bool use24HourClock = ClockIdentifier == "24HourClock";

            var columnsD = use24HourClock ? "*, Auto, *" : "*, Auto, *, Auto, *";
            _contentGrid.ColumnDefinitions = new ColumnDefinitions(columnsD);

            _thirdPickerHost!.IsVisible = !use24HourClock;
            _secondSplitter!.IsVisible = !use24HourClock;

            Grid.SetColumn(_firstPickerHost!, 0);
            Grid.SetColumn(_secondPickerHost!, 2);

            Grid.SetColumn(_thirdPickerHost, use24HourClock ? 0 : 4);

            Grid.SetColumn(_firstSplitter!, 1);
            Grid.SetColumn(_secondSplitter, use24HourClock ? 0 : 3);
        }

        private void SetSelectedTimeText()
        {
            if (_hourText == null || _minuteText == null || _periodText == null)
                return;

            var time = SelectedTime;
            if (time.HasValue)
            {
                var newTime = SelectedTime!.Value;

                if (ClockIdentifier == "12HourClock")
                {
                    var hr = newTime.Hours;
                    hr = hr > 12 ? hr - 12 : hr == 0 ? 12 : hr;
                    newTime = new TimeSpan(hr, newTime.Minutes, 0);
                }

                _hourText.Text = newTime.ToString("%h");
                _minuteText.Text = newTime.ToString("mm");
                PseudoClasses.Set(":hasnotime", false);

                _periodText.Text = time.Value.Hours >= 12 ? CultureInfo.CurrentCulture.DateTimeFormat.PMDesignator :
                    CultureInfo.CurrentCulture.DateTimeFormat.AMDesignator;
            }
            else
            {
                _hourText.Text = "hour";
                _minuteText.Text = "minute";
                PseudoClasses.Set(":hasnotime", true);

                _periodText.Text = DateTime.Now.Hour >= 12 ? CultureInfo.CurrentCulture.DateTimeFormat.PMDesignator :
                    CultureInfo.CurrentCulture.DateTimeFormat.AMDesignator;
            }
        }

        protected virtual void OnSelectedTimeChanged(TimeSpan? oldTime, TimeSpan? newTime)
        {
            SelectedTimeChanged?.Invoke(this, new TimePickerSelectedValueChangedEventArgs(oldTime, newTime));
        }

        private void OnFlyoutButtonClicked(object? sender, Interactivity.RoutedEventArgs e)
        {
            if (_presenter == null)
                throw new InvalidOperationException("No DatePickerPresenter found.");
            if (_popup == null)
                throw new InvalidOperationException("No Popup found.");

            _presenter.Time = SelectedTime ?? DateTime.Now.TimeOfDay;

            _popup.PlacementMode = PlacementMode.AnchorAndGravity;
            _popup.PlacementAnchor = Primitives.PopupPositioning.PopupAnchor.Bottom;
            _popup.PlacementGravity = Primitives.PopupPositioning.PopupGravity.Bottom;
            _popup.PlacementConstraintAdjustment = Primitives.PopupPositioning.PopupPositionerConstraintAdjustment.SlideY;
            _popup.IsOpen = true;

            // Overlay popup hosts won't get measured until the next layout pass, but we need the
            // template to be applied to `_presenter` now. Detect this case and force a layout pass.
            if (!_presenter.IsMeasureValid)
                (VisualRoot as ILayoutRoot)?.LayoutManager?.ExecuteInitialLayoutPass();

            var deltaY = _presenter.GetOffsetForPopup();

            // The extra 5 px I think is related to default popup placement behavior
            _popup.VerticalOffset = deltaY + 5;
        }

        private void OnDismissPicker(object? sender, EventArgs e)
        {
            _popup!.Close();
            Focus();
        }

        private void OnConfirmed(object? sender, EventArgs e)
        {
            _popup!.Close();
            SelectedTime = _presenter!.Time;
        }
    }
}

redpajama_set_name: RedPajamaGithub


text:
Egypt Tourism Minister invites Oprah
Egypt's new Minister for Tourism, Munir Fakhry Abdel Nour, has invited Oprah Winfrey to visit Egypt in an effort to revive tourism in the country. After the uprising in Egypt which ousted President Husni Mubarak, the new government is trying to restore normalcy in the country. The newly appointed Tourism Minister has invited Oprah as well as other international celebrities to visit Egypt. Concerts and events are being planned to take place in Tahrir Square which was earlier the scene of violent protests by pro-dmocracy forces in Egypt.
Tourism accounts for a major share of Egypt's economy and reviving it is a high priority. A large number of people work in the travel sector and have been badly affected by the decine in tourism. Many European tourists have changed their travel plans and are now going to Spain and the Canary Islands. Egypt expects that by inviting celebrities it will be able to give a clear message to tourists that the country is once again safe for travel.
The US talk show queen, Oprah Winfrey, hosted her show in Australia in December and attracted a tremendous amount of interest in travel to Australia.
Glass and Ceramics Museum
located at No. 75, 30 Tir St, in the Tajrish Sq Area, this interesting collection is extremely well laid out and displayed. The shop on the ground floor sells a guidebook to the exhibits. The collections are varied and eclectic and the buildings themselves are quite beautiful. It was built in the time of the Qajar Shahs as a private residence and was previously used as an embassy for Egypt. ...
Livrustkammaren
Situated in the vaults underneath the Royal Palace, The Royal Armory Museum is truly a historical treat. The museum's collection spans over 500 years of royal history replete with sagas of love and murder, weddings and death, victory and defeat. While at the museum, you can enjoy everything, from royal costumes, weapons, armors and carriages to preserved stomach contents of conspirators and assass...
Parque Los Caobos
Caracas, Venezuela
Address: east of the Parque Central Complex One of the oldest parks in Caracas, the Parque Los Caobos is known widely for its enchanting mahogany trees from which the park derives its name. A walk through the park, on its gently winding trails lined with hedges in fresh bloom, is a relaxing way to let go of the day's troubles and rejuvenate your mind and body for the next day. The park is us...
What to do with Children in Tow
Kids have many things to see and do when in Amsterdam: ARTIS ZOO Founded in 1838, the Artis Zoo is one of the largest in Europe. It is home to 700 animal species and 200 tree varieties, many on the verge of extinction. Artis is also one of the fewest zoos in Europe that has famous historical buildings [the Large Museum (1855), the unique Library building (1867) and the Aquari...
Cruise into the sunset on your own boat...

redpajama_set_name: RedPajamaCommonCrawl


text:
'use strict';

const libs = {
  sodium: sodium => ({
    open: sodium.api.crypto_secretbox_open_easy,
    close: sodium.api.crypto_secretbox_easy,
    random: n => sodium.randombytes_buf(n),
  }),
  'libsodium-wrappers': sodium => ({
    open: sodium.crypto_secretbox_open_easy,
    close: sodium.crypto_secretbox_easy,
    random: n => sodium.randombytes_buf(n),
  }),
  tweetnacl: tweetnacl => ({
    open: tweetnacl.secretbox.open,
    close: tweetnacl.secretbox,
    random: n => tweetnacl.randomBytes(n),
  }),
};

exports.methods = {};

(async () => {
  for (const libName of Object.keys(libs)) {
    try {
      const lib = require(libName);
      if (libName === 'libsodium-wrappers' && lib.ready) await lib.ready; // eslint-disable-line no-await-in-loop
      exports.methods = libs[libName](lib);
      break;
    } catch (err) {} // eslint-disable-line no-empty
  }
})();


text:
EIA: Natural gas topped coal for fifth consecutive month in November
Flickr; Tim Evanson
Robert Walton @TeamWetDog
Natural gas-fired generation was on pace for a strong finish in 2015, and though data for December are still pending, the U.S. Energy Information Adminsitration says it exceeded coal generation in at least six months of the year.
Before April 2015, natural gas' share of the United States generation mix had never exceeded coal, but according to EIA's "Electric Power Monthly" report, natural gas led coal in November, making it the fifth consecutive month and sixth time overall it has accomplished this feat in 2015 thus far, SNL Energy reports.
Coal generation is still likely to top natural gas for full-year 2015, but in November EIA reports that coal generated 87,789 GWh to natural gas' 101,866 GWh.
Coal-fired generation will still likely be king in 2015, but the gap has been closing and gas appears to have finished the year with a remarkable run. Coal generation tumbled nearly 10% between October and November, pressured by a mild winter, SNL Energy reports. While gas generation dipped as well, the decline was not as significant.
Coal generated 87,789 GWh in November, compared with 97,409 GWh in October. Gas generated 101,866 GWh, compared with 109,921 GWh in October. Nuclear generation remained roughly constant over the two months, supplying more than 60,000 GWh.
Before 2015, natural gas generation had never surpassed coal, but it did so last year in April, July, August, September, October and, now, November. Data for December will be available in about a month. Overall generation has fallen since the summer, declining from more than 400,000 GWh in July to about 300,000 GWh in November.
There are indications the U.S. is moving away from coal. EIA said coal production last year was estimated to decline by 92 MMst, with the production declines occurring in all regions (led by the Appalachian region, with a 12% production decline). And carbon regulations under the Clean Power Plan, finalized last year, would make it nearly impossible to build new coal generation facilities without expensive pollution controls if the rules are not overturned in court.
SNL Energy EIA: Gas tops coal generation for 6th time in first 11 months of 2015
Follow Robert Walton on Twitter
Filed Under: Generation


text:
Paul Gallagher
pgallagher@hausfeld.com Email
Paul brings a breadth of litigation experience that is rare to find in this age of legal specialization. For eight years prior to joining Hausfeld, Paul was a white-collar prosecutor with the Antitrust Division of the United States Department of Justice in Washington, D.C., specializing in prosecution of criminal antitrust cases against corporations and executives engaged in illegal activity globally and within the United States.
Paul has spearheaded some of the largest and most complicated class actions ever litigated in the United States across a wide range of subject matters, including antitrust, civil rights, consumer rights, human rights, and RICO claims. He consistently has been involved in cases on the cutting-edge of the law, raising and pursuing novel legal theories and arguments that have expanded the law and claimants' access to justice.
While at the Department of Justice, Paul was heavily involved in the Antitrust Division's investigation and prosecution of cartels in the global automotive industry, an investigation that resulted in charges against over 110 corporations and executives and the recovery of almost $3 billion in criminal fines. The investigation was the largest in the Antitrust Division's history.
Before joining the Justice Department, Paul was the Chief of the Public Advocacy Section of the District of Columbia Attorney General's Office, where he supervised the investigation and prosecution of civil cases involving consumer fraud, antitrust, qui tam and other litigation on behalf of the residents of the District of Columbia. He recovered millions of dollars in fines and restitution on behalf of D.C. residents.
Empagran, S.A. et al. v. F. Hoffmann-LaRoche, Ltd. - Paul argued this case in the D.C. Circuit Court of Appeals on behalf of a class of foreign vitamin purchasers, which was designated as the "Matter of the Year" by the Global Competition Review. The case was ultimately heard by the U.S. Supreme Court, resulting in a groundbreaking decision clarifying the scope of U.S. Sherman Act jurisdiction.
McLaughlin v. Philip Morris – Paul was one of the lead counsel in this historic consumer-RICO class action litigation on behalf of cigarette smokers who were defrauded by the major global cigarette manufacturers into purchasing "light" cigarettes by representations that those cigarettes were less harmful than other cigarettes. At the time, the case was the largest class action ever certified in a U.S. court.
In re Swiss Banks – Counsel in an action against both the Swiss government and the major banks in Switzerland for their illegal receipt, liquidation and retention of assets owned by Holocaust victims and their heirs, which resulted in recovery of over $1.2 billion for class members
Paul was one of the principal counsel who prosecuted a series of human rights cases that resulted in the recovery of damages resulting from certain countries' and companies' violations of customary international law during World War II.
In re German Slave and Forced Labor – Counsel in a class action against the German government and a wide spectrum of German industry for their use of slave and forced labor during World War II, which resulted in the establishment of a victims' fund of $6.5 billion dollars.
During his time with the Department of Justice, Paul received numerous awards, including the Attorney General Distinguished Service Awards and the Health and Human Service Inspector General's Awards for Fighting Fraud.
William & Mary Law School, J.D.; Member, William & Mary Law Review, 1992
George Mason University, B.A. Government and Politics, 1986
Related News & Press
Hausfeld Adds Veteran DOJ Litigator Paul Gallagher to Washington, D.C. Office


This article is the result of an interview with David Capozzi, Executive Director at the US Access Board.
On January 18, 2017, the U.S. Access Board published a final rule that updates accessibility requirements for information and communication technology (ICT) in the federal sector covered by Section 508 of the Rehabilitation Act. The rule also updates guidelines for telecommunications equipment subject to Section 255 of the Communications Act.
The Access Board issued the original 255 Guidelines for telecommunications equipment and customer premises equipment in 1998. Two years later, in 2000, the Board published the original 508 Standards. The underlying laws (Section 508 of the Rehabilitation Act and Section 255 of the Communications Act) require the Board to periodically review and, as appropriate, amend the standards and guidelines to reflect technological advances or changes in technology. This is the first update of the original standards and guidelines.
The 2017 final rule was based on input from the Board's Telecommunications and Electronic and Information Technology Advisory Committee. The committee was comprised of 41 members from a broad cross-section of stakeholders representing industry, disability groups, and government agencies. The committee also included international representatives from the European Commission, Canada, Australia, and Japan. The Board also received feedback from two advance notices of proposed rulemaking in 2010 and 2011 and a proposed rule in 2015. In total, we received 637 public comments and held 7 public hearings.
Technological advances over the past two decades have resulted in the widespread use of multifunction devices that called into question the ongoing utility of the product-by-product approach used in the Board's original 508 Standards and 255 Guidelines. Consequently, one of the primary purposes of the final rule was to replace the product-based approach with requirements based on functionality, and, thereby, ensure that accessibility for people with disabilities keeps pace with advances in ICT.
The Revised 508 Standards and 255 Guidelines incorporate by reference the Web Content Accessibility Guidelines (WCAG) 2.0, a globally-recognized and technologically-neutral set of accessibility guidelines for Web content. For Section 508-covered ICT, all covered Web and non-Web content and software – including, for example, Web sites, intranets, word processing documents, portable document format documents, and project management software – is required, with a few specific exceptions, to conform to WCAG 2.0's Level A and Level AA Success Criteria and Conformance Requirements. By applying a single set of requirements to Web sites, electronic documents, and software, the revised requirements adapt the existing 508 Standards to reflect the newer multifunction technologies (e.g., smartphones that have telecommunications functions, video cameras, and computer-like data processing capabilities) and address the accessibility challenges that these technologies pose for individuals with disabilities.
The final rule harmonizes its updated requirements with other guidelines and standards both in the U.S. and abroad, including EN301549 "Accessibility requirements suitable for public procurement of ICT products and services in Europe" and with WCAG 2.0. Throughout our update process the Board worked very hard to ensure consistency with other consensus guidelines and international standards to promote global harmonization and facilitate compliance. ICT requirements that are closely aligned remove ambiguity, increase marketplace competition, and lead to better accessibility features and outcomes.
The Revised 508 Standards specify that all types of public-facing content, as well as nine categories of non-public-facing content that communicate agency official business, have to be accessible, with "content" encompassing all forms of electronic information and data. The existing standards require Federal agencies to make electronic information and data accessible, but do not delineate clearly the scope of covered information and data. As a result, document accessibility has been inconsistent across Federal agencies. By focusing on public-facing content and certain types of agency official communications that are not public facing, the revised requirements bring needed clarity to the scope of electronic content covered by the 508 Standards and, thereby, help Federal agencies make electronic content accessible more consistently.
Federal agencies will have until January 2018 to comply with the Revised 508 Standards. This extended period for compliance is responsive to some agencies' concerns about the time it will take them to make ICT compliant with the Revised 508 Standards. In addition, the Revised 508 Standards include a "safe harbor" provision for existing (i.e., legacy) ICT. Under this safe harbor, unaltered, existing ICT (including content) that complies with the original 508 Standards need not be modified or upgraded to conform to the Revised 508 Standards. This safe harbor applies on an element-by-element basis in that each component or portion of existing ICT is assessed separately.
Every two years the Department of Justice is required to issue a report containing information and recommendations regarding the extent to which electronic and information technology of the Federal Government is accessible to and usable by individuals with disabilities. The latest report was issued in September 2012.
Slightly more than 50% of agency components had established a general policy to implement and comply with Section 508.
Nearly 75% of agency components had appointed a Section 508 coordinator, but only 35% of agency components had established a Section 508 office or program.
Agency components experienced difficulty in providing Section 508 training to their personnel - only 40% of agency components provided Section 508 training, and these agency components provided only a small number of hours of training to their agency acquisition workforce (less than one hour) or ICT developers (a little more than one hour).
A majority of the agency components currently incorporate accessibility provisions into their ICT procurements. However, less than 50% of agency components incorporated specific applicable Section 508 Standards as requirements in each procurement solicitations. A majority of the agency components only provided standardized compliance language or clauses in their solicitations. Additionally, instead of relying on actual product testing to validate Section 508 compliance, agency components most often relied on reviewing the materials submitted by the contractor or vendor to evaluate whether the deliverables met the Section 508 requirements.
Agency components reported 140 administrative complaints and seven civil actions since June 21, 2001, the effective date of the Section 508 Standards. However, most of the administrative complaints and civil actions were filed against components in very large agencies.
70% of agency components reported having accessibility policies in place for website development and that their web pages, web-based forms, and web-based applications were generally accessible. Also, nearly 58% of agency components reported performing routine automated and/or manual evaluation and remediation on their websites. Agency components reported some difficulty, however, with providing captioning and audio description for multimedia content and providing keyboard accessibility.
The Board conducted two webinars on the final rule on January 31 and February 2. The webinars provided an overview of the final rule's scope and structure, highlighted substantive changes from earlier requirements, and answered questions from attendees.
On March 2, 2017, the Access Board postponed by one day the effective date of its final rule updating accessibility requirements for information and communication technology (ICT) covered by Section 508 of the Rehabilitation Act and Section 255 of the Communications Act which was published on January 18. This action was taken in response to a White House memorandum that directs federal agencies to delay for 60 days the effective date of recently published rules that, as of the date of the memorandum (January 20), had not yet taken effect. In keeping with the White House memorandum, the Access Board has postponed the effective date of the ICT final rule until March 21. This amounts to a one-day delay in the effective date of the rule relative to its originally-published effective date (March 20), as indicated in a notice the Access Board published in the Federal Register.
The postponed effective date does not change the scoping or technical requirements in the updated 508 Standards or 255 Guidelines. Nor does it alter compliance dates of the rule. Compliance with the updated Section 508 Standards is not required until January 18, 2018 or, in the case of ICT procurements, dates to be established by the Federal Acquisition Regulatory Council. The compliance date for the Section 255 Guidelines will be set by Federal Communications Commission when it adopts the guidelines.


In the venue Allaktivitetshuset in Sundbyberg, in our home town we arranged the event "Kropp & Själ" in an old cinema theatre that has been remade for events and meetings for locals. Together with our child choir Rising Star we have been practising singing and dancing during the winter and did a splendid show of performances with our own created songs that were released recently as well as some cover songs with simple moves to get the audience going. After all the theme was HEALTH so we wanted all to get their bodies in action in a light and happy way. All ages were welcome for free so the place was crowded with joyful people interested to watch performances and get to know more about how they could improve their health.
On this sunny spring day the guests were encouraged to dress in GREEN, take press photos on the green carpet and were invited to try fresh drinks and healthy snacks.
Sara Ekholm is a life coach and medium working with healing who presented her work and told us about her life story. She was definitely the highlight of the evening acting as conferencier during the whole event. What a woman!
Charlotta Berglund showed us how to do yoga, presented bemer and is also a medium, doing massage and animal consulting.
Shani Panabokke showed acupunture and ayurveda and her daughter Dilini gave us some lovely singing on stage. Karina Tyglare was talking about her work treating mental and physical illness and Anette Blom Thomasson presented healing and beauty for skin and spa.
We preciously hosted several appreciated shows with the choir in Allaktivitetshuset through our registered association Rising Star aimed for music activities with youths up to 20 years old. We received a scholarship in 2017 for our achievements as song coaches and for the cultural life of Sundbyberg. During the last four years we have worked our way up from rehearsing in a class room to turning the school aula into a decorated stage for the whole school to be our audience.
We had as many as 27 girls in the choir Rising Star from class 1-5 who have all become very confident on live performances on stage and in front of the camera. We love to be their inspiration and see them grow, but being all the roles of the song coach, stage coach, songwriter, event arranger and organizer, music and video producer, sound technician and running a company eventually also takes away the spark we once had, usually having a lot of energy to start new projects. We feel now more than ever that we need more support in the future from our followers, coworkers and to keep running the same pace! We are grateful to the people who are always there for us in this crazy music business!
Please follow our Youtube channel, Facebook and Instagram for follow back.


So why is Verizon overpaying by roughly 30 billion dollars? Who knows? Perhaps some of the board members, executives, or large shareholders of Verizon are also shareholders of Vodafone. Good for those folks, but not for the remaining shareholders.
This entry was posted in Financial, Technology. Bookmark the permalink.


Consider booking a private bus trip for your group or organization on our luxury motorcoaches. In doing so, you can feel comfortable knowing that our staff – from our bus drivers and mechanics to our professional staff – keeps your satisfaction and safety at the forefront of our business.
Whether you are looking to book a charter for a short trip to New York City, Boston, Montreal, or a motorcoach for long-distance trip across the country, you can rely on us to provide professional, reliable service. No trip or event is too small or too far.
Our commitment to excellence has made us the most respected bus transportation provider in Upstate New York. So, sit back and relax, we'll take care of everything for you!


Clinical Engineering Services (CES) is a system-wide clinical equipment maintenance and technology assessment service operated as a department of SSM Integrated Health Technologies (IHT).
Today's modern medicine relies heavily upon "State of the Art" technology, which must be properly maintained to ensure reliable and safe operation when called upon. CES' inhouse staff of 120 clinical engineers provides an efficient and cost effective alternative to expensive outsourcing. It is estimated that CES saves SSMHC approximately $11 million annually in clinical equipment maintenance expenses compared with outsourcing.
CES also provides a leading role in the evaluation and negotiations for new clinical devices to ensure SSMHC purchases the best devices at the lowest cost. Today, CES manages the clinical engineering programs in 15 wholly owned SSMHC hospitals, as well as over a dozen non-SSM or partially owned facilities (outreach). We are responsible for the maintenance and management of approximately 65,000 clinical devices.
CES has two principle goals supporting our SSMHC mission, "Through Our Exceptional Health Care Services, We Reveal the Healing Presence of God:"
1. Provide the highest standard of maintenance, equipment management, and technology assessment.
2. Reduce and control clinical equipment service costs.


WELCOME TO THE MUMMY MAKE-UP PROJECT! I'M FIVE YEARS SHY OF 40 AND AFTER 20 YEARS OF CLUELESS MAKE-UP APPLICATION, 2015 IS THE YEAR OF TURNING DRAB TO FAB. I'M GOING TO FIND THE BEST SKINCARE AND MAKE-UP PRODUCTS, SHARE THEM WITH YOU AND ACTUALLY LEARN HOW TO USE THEM. I WOULD LOVE YOUR ADVICE ALONG THE WAY AS WELL AS YOUR SUPPORT – I'M SURE I'M NOT THE ONLY MUMMY OUT THERE WHO IS COMPLETELY CLUELESS ABOUT MAKE-UP!
So, in keeping with my new resolution to actually spend a little bit of time each day taking care of my skin, I have added a product to my limited routine which can get all of it's work done at night while I'm catching up on (not enough!) sleep.
You won't be surprised to hear that I have never used a night cream before. Previously, if I bothered to remove my makeup before bed, I would just pop on whatever daily moisturizer I was currently using and call it a day.
Sadly, advancing age and a recurring failure to drink enough water means I often wake with my skin feeling dry and looking sallow and worse than it did before bed!
Some new products take longer to get used to than others, as such, it's taken about a week for my skin to settle down after introducing Trilogy Rosapene Night Cream. This is an intense moisture surge that initially left my skin feeling slightly oily and caused a few spots. Happy to report that this is now a thing of the past and my skin looks positively freaking dewy when I wake up! Adding a night cream has made a huge difference in how my skin feels throughout the day, starting with a hydrated base is resulting in a much more natural, smooth makeup application, and I find that afternoon oil-slicks are now a thing of the past. I'm picking this is probably due to my skin no longer feeling like it's got to produce huge amounts of oil to compensate for it's dehydrated state?
Water loss and blood flow are highest at night ergo; the skin is most receptive to nourishing natural activities while we sleep.
I was gifted this product for editorial consideration. All opinions expressed are my own.


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ICCSS paid tribute to Polish officers buried at the Polish War Cemetery in Kharkiv
ICCSS strengthens cooperation with Ukraine on industrial cybersecurity
Honorary President of the German Federal Agency for Technical Relief joined ICCSS Global Team
ICCSS Global Team enhances worldwide chemical and environmental safety and security, civil protection and cybersecurity
President of the German Federal Agency for Technical Relief visited ICCSS
ICCSS
CHEMSS
International Centre for Chemical Safety and Security
About ICCSS
ICCSS Leaders
ICCSS in the World
Chemical safety and Security in Jordan and the Middle East
EU-CHEM-REACT
EU-CHEM-REACT-2
Chemical Security in Ukraine
Kenyan Program for Chemical Safety & Security
Partnership in Horizon 2020
Local Awareness and Responsibility
Tailor-made trainings
TableTop Methodology
Chemical security manual for Trainers
Working together to enhance
chemical safety and security
Go to news | Our global approach
International Days on Chemical Safety and Security and Cybersecurity
Warsaw, Vistula University, 9-10 December 2019
About International Days
Join ICCSS global projects
CHEMSS | Our projects
ICCSS unique methodology and training courses:
Tabletop methodology | Training programmes
The ICCSS President, Amb. Krzysztof Paturej and Chancellor of Vistula University, Dr. Zdzislaw Rapacki and their Ukraine partners from the National...
The ICCSS President, Amb. Krzysztof Paturej and Chancellor of Vistula University, Dr. Zdzislaw Rapacki visited on 08-09 January, 2020, the National...
Tweets by krispaturej Tweets by ICCSS_PL
Want to organize chemical security training but you need additional funding? Submit your application to CSP!
CSP - Chemical Security Program - is a project sponsored by the US Department of State. Since its inception in 2007, CSP has supported a variety of activities to promote chemical security in partner countries, including: chemical safety and security trainings for academic and industry audiences, Organisation for the Prohibition of Chemical Weapons (OPCW)-led chemical security workshops for government and industry representatives, trainings to strengthen academic chemical societies, the development of a chemical security toolkit, architectural design support for chemistry lab construction, agrichemical security initiatives, and grants for training and physical security upgrades at academic and industrial facilities with dangerous chemicals.
To see Funding opportunities, click here.
The ICCSS promotes the chemical safety and security culture by bringing the issue to national level, by being more efficient in chemical safety and security capacity building and best practices exchanges, and by improving national and international coordination of chemical safety and security actors. Based on these core considerations the ICCSS has introduced five leading principles: Sustainability; Continuity; Public-private partnership; Multi-stakeholder participation; and Modern management.
The International Centre for Chemical Safety and Security (ICCSS) serves as a centre of excellence in chemical safety and security and supports the development of national and international networks of chemical safety and security officers from relevant national agencies, international organizations, industries, and civil society.
The ICCSS provides continuity to the international efforts in chemical safety and security and focuses on building the potential and expanding capabilities for research, development, storage, production, and safe use of chemicals for peaceful purposes.
The ICCSS serves as a multi-stakeholder chemical safety and security forum for exploring sustainable ways and means of sharing knowledge and best practices, promoting public-private enterprises and modern management in all areas of chemical activities:
industry;
transportation of chemicals;
academic activities and studies;
scientific research; and
laboratory work.
The ICCSS combines and multiplies efforts, technical resources and expertise from industry associations, international organizations, national agencies, and NGOs, to achieve chemical safety and security in all aspects/areas of chemical activities.
PartnersAll Partners
The ICCSS currently cooperates with a number of national and international partners in a variety of activities and capacities, inter alia:
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The International Centre for Chemical Safety and Security serves as a centre of training in chemical safety and security and supports the development of national and international networks of chemical safety and security officers from relevant international organisations.
ul. Leszno 8/1, 01-192 Warszawa
Wdrożenie TYPO3 - Data Quest


Losing weight is very difficult. More people are overweight than ever before. Due to technological advancements that have resulted in sedentary lifestyles and a proliferation of fast food restaurants, it seems like just about everyone has felt the need to lose weight at one time or another. If you are sick and tired of being overweight and are ready to get into the best shape of your life, this is the perfect article for you. Read on to learn several terrific weight loss tips. No matter how overweight you are, if you are serious and make a real commitment you can achieve all of your weight loss goals and have the type of body that you have always dreamed about.
It is impossible to lose weight without following a healthy, reduced calorie diet. There is truth to the old saying "you are what you eat." If you eat high calorie, fatty foods, you will never be slim and trim. Your daily diet should consist of primarily vegetables, whole grains and lean proteins, such as chicken breasts, egg whites and tuna. Avoid sugary sweets and unhealthy fast foods. Stay away from simple, fast digesting carbohydrates such as white bread. Your diet should be low in unhealthy saturated fats, but not completely fat free. The unsaturated fats found in certain fatty fish, nuts and certain oils can be very beneficial to your health.
Along with monitoring the foods you eat, pay careful attention to the beverages that you drink. Fruit juices and soda pop contain tons of empty, unhealthy calories. Drink 6 to 8 glasses of water every day, and you may be surprised at how much weight you lose. Water is completely free of calories and drinking a lot of it will keep your from feeling hungry.
To lose weight you must be active. There is no way around it, in order to lose weight you need to engage in regular exercise. Strive to perform some type of aerobic exercise 4 to 6 days per week. You can sign up at a gym and take an exercise class if you would like or you could simply go for long walks or bicycle rides. Even daily chores, such as yard work and cleaning will burn off calories. Get up and move!
Studies have shown that dieters who keep a daily journal are much more likely to reach their weight loss goals compared to those who do not. Write down the foods you eat, the exercises you perform and any thoughts or feelings you are having. Weigh yourself once per week and track your weight loss progress in your journal.
Find a weight loss partner. Having a friend join you on your weight loss journey can make the experience much more enjoyable. In addition to making exercise and dieting more tolerable, a friend can lift up your spirits when you are feeling discouraged.
Weight loss is not easy for everyone, but with hard work, patience and dedication you can reach all of your goals. Remember, never give up on yourself!


A Lower Salford man will spend time in state prison for attacking his wife and her friend.
Gary A. Eder, 45, of the 200 block of Jan Drive, on Wednesday received a 1�- to three-year state prison sentence and a 10-year probation sentence after pleading guilty in Montgomery County Court to felony charges of burglary and aggravated assault.
The break-in and subsequent assault occurred on Aug. 24, 2013, at a residence in the 400 block of Rutledge Court in Montgomery Township, according to court documents.
Eder�s wife, who shared the Lower Salford home with her husband, told police she had gone to stay at a friend�s house because of her constant fights with Eder, according to the criminal complaint.
On the day of the assault, Eder�s wife said she had received repeated phone calls from him, threatening her and a male friend, according to the complaint.
Later that day, Eder forced open the door of the residence where his wife was staying and began hitting her and her friend, the complaint said. Police said in the complaint that the male friend had a cut above his eye and his eye was swollen shut and the woman's nose was swollen.


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Shoaib Mansoor's 'Verna' unveils its trailer
Verna, the latest offering by Shoaib Mansoor is ready to hit the cinemas on 17th November, 2017 worldwide. The special trailer launch of the film was hosted on 17th October at the Neuplex cinema in Karachi where the leads of the movie, Mahira Khan and Haroon Shahid, were present, and unveiled the trailer to the members of the media for the first time.
Verna is written, produced and directed by Shoaib Mansoor, known for his highly acclaimed films Khuda Ke Liye and Bol. The rest of the cast includes Zarrar Khan, Rasheed Naz and Naimal Khawar.
Speaking at the trailer launch, Mahira Khan stated, "I was overwhelmed to receive such a splendid response to the first teaser and song. The whole team has put a lot of effort in this movie and I hope this movie leaves an impact on people."
"It's an honor for me to debut through Shoaib Mansoor's movie. He has always believed in giving fresh talent a chance and I feel lucky to be a part of this movie. I cannot wait for the movie to be released and get audience's feedback," stated Coke Studio sensation, Haroon Shahid, who was present at the trailer launch.
Shoaib Mansoor, the man behind Verna, stated, "Film to me is an art form like poetry. Many people may not agree with me, but I believe in giving art some meaning. All over the world, poetry done with some deeper meanings and philosophy has always lived longer. Whether my films pass for being artistic may rightly be questionable, but they do reflect my beliefs and conscience. Things happening around me bother and hence, I am forced by my inner self to take them on. My films reflects the anger that I feel on social injustice of all forms. So every time I am about to embark on a film it is my anger that controls my thoughts, not the hunger for success and money."
"That doesn't mean that I have a thing against films based on pure entertainment. Not at all. By all means they should be made and succeed, and I have all the due respect for their makers. I am happy at the recent success of Punjab Nahi Jaungi and Na Maloom Afraad and wish success to all others that are in line to hit the screen. We need those films to rejuvenate the industry in our country, but we also need some to say something thought provoking. I hope I am one of those." he further added.
The story of Verna revolves around the lives of a highly educated and beautiful Sara (Mahira Khan) and Aami (Haroon Shahid). The young couple lives a splendidly happy life until their lives falls apart due to an earth shattering incident in their life. Verna portrays the story of a strong women who doesn't believe in being a victim, but decides to fight for herself against all odds. It addresses tabooed subjects circling around a woman's honor and the most commonly held myths and biases around rape.
The film will be distributed under the banner of HUM Films, and Sultana Siddiqui, the president of HUM Network was present at the trailer launch. She stated, "The uniqueness in Shoaib Mansoor's movies lies in the fact that they revolve around social issues, and aim to educate the audiences. I am simply amazed to see the strong impact that the trailer has left on audiences."
Listen to all VERNA songs right here on Taazi.com
http://taazi.com/Verna
#HaroonShahid 1#HumFilms 1#MahiraKhan 1#ShoaibMansoor 1#Verna 2
Here's a beautiful message by Ali Zafar to Mahira Khan!
Ahat Si' – Another mystical soundtrack by Abida Parveen releasing exclusively with Taazi!
Verna's Music Released


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Alfred Karl Neuland (ur. 10 października 1895 w Valdze, zm. 16 listopada 1966 w Tallinnie) – estoński sztangista, dwukrotny medalista olimpijski, a także mistrz świata.

Kariera
Reprezentował klub Kalev Tallinn, w swoich pierwszych zawodach uczestniczył w wieku 16 lat w Rydze, w 1911 roku. W 1915 roku został mistrzem Rosji w wadze lekkiej, a rok później był najlepszy w wadze średniej. Walczył w I wojnie światowej, a po jej zakończeniu w wojnie estońsko-bolszewickiej.

Pierwszy sukces w karierze osiągnął w 1920 roku, kiedy podczas igrzysk olimpijskich w Antwerpii zdobył złoty medal w wadze lekkiej. W zawodach tych pokonał dwóch Belgów: Louisa Williqueta i Florimonda Roomsa. Na rozgrywanych dwa lata później mistrzostwach świata w Tallinnie także zwyciężył, wyprzedzając dwóch rodaków: Eduarda Vanaasemego i Voldemara Noormägiego. Wystartował także na igrzyskach olimpijskich w Paryżu, gdzie wywalczył srebrny medal w wadze średniej. Rozdzielił tam na podium Włocha Carla Galimbertiego i kolejnego Estończyka, Jaana Kikkasa.

Neuland był pierwszym mistrzem i multimedalistą olimpijskim reprezentującym Estonię. W latach 1920–1925 sześciokrotnie ustanawiał rekordy świata w kategoriach lekkiej i średniej.

Po zakończeniu kariery sportowej został przedsiębiorcą; pracował także jako trener i sędzia sportowy. W 1995 roku postawiono mu pomnik w Valdze.

Wyniki

Przypisy

Linki zewnętrzne

Estońscy medaliści olimpijscy
Estońscy sztangiści
Medaliści Letnich Igrzysk Olimpijskich 1920
Medaliści Letnich Igrzysk Olimpijskich 1924
Urodzeni w 1895
Zmarli w 1966
Ludzie urodzeni w Valdze

redpajama_set_name: RedPajamaWikipedia


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Manchester United showed "big heart and soul" to twice come from behind to earn a draw against Arsenal, said manager Jose Mourinho.
Mourinho dropped high-profile duo Paul Pogba and Romelu Lukaku after recent disappointing performances which have brought only three points from the past 12 available – and, while quality was lacking, no-one could doubt his side's endeavour and character.
Goalkeeper David de Gea made an uncharacteristic error as he failed to deal with Shkodran Mustafi's tame 26th-minute header, which Ander Herrera could not clear before it crossed the line.
United were swiftly level through Anthony Martial after Arsenal keeper Bernd Leno could only half-clear a free-kick to set up an exciting second half.
Substitute Alexandre Lacazette scrambled Arsenal back in front after 68 minutes but United were on terms once more within seconds as Jesse Lingard pounced on Sead Kolasinac's attempted backpass.
De Gea redeemed himself with a succession of fine saves as both sides had to content themselves with a point, with United now eighth in the Premier League and Arsenal, who extended their unbeaten run this season to 20 games, dropping to fifth.
"As always we made mistakes and we paid for the mistakes," said Mourinho.
Mourinho demonstrated he would not shy away from taking the big decisions as Pogba and Lukaku were consigned to the bench – he will have known the spotlight would have shone on him in unforgiving fashion had United lost.
Elsewhere Wolves boss Nuno Espirito Santo challenged his side to "maintain these standards" after they fought back from a goal down to stun Chelsea and gain their first victory in seven Premier League matches.
Two goals in four second-half minutes – from Raul Jimenez and Diogo Jota – gave the hosts victory after Ruben Loftus-Cheek's 25-yard effort was headed into his own net by Wolves captain Conor Coady after only 18 minutes.
"This doesn't mean all the problems are solved," said Nuno, whose side won for the first time since their 1-0 away victory at Crystal Palace on 6 October.
"It was a very good performance. In the first half, the organisation and staying in the game was important.
Wolves' win leaves them 12th in the Premier League, only four points behind sixth-placed Everton.
And at Turf Moor, Substitute Roberto Firmino scored with his first touch as Liverpool came from behind to beat struggling Burnley and stay in touch with leaders Manchester City in the Premier League table.
Jack Cork gave the hosts a surprise lead after Alisson could not gather the ball following a corner but Jurgen Klopp's side hit back with three goals.
James Milner equalised with a superb low finish from the edge of the box and Firmino, one of seven players left out from the side that started against Everton on Sunday, tapped home Virgil van Dijk's athletic cutback.
Xherdan Shaqiri made sure of the win for Liverpool with a clinical counter-attack goal moments after Alisson had saved superbly from Ben Mee.
Liverpool are two points behind Pep Guardiola's side after 15 games and have made the best start to a season in the club's 126-year history.
They will go top of the table and put pressure on City on Saturday if they win at Bournemouth (12:30 GMT), with the leaders facing a difficult game at Chelsea at 17:30.


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Some Oscar suspense remains despite 'No Country,' 'Blood' momentum
Feb 24, 2008 at 12:01 AM Jul 1, 2009 at 12:56 PM
LOS ANGELES - As usual in Hollywood, Oscar night looks like the same people will generally be collecting the same prizes. Yet some suspense remains for a few of the main awards. A look at the key categories:
The Coen brothers' "No Country for Old Men" is the odds-on favorite to claim best picture, winning top honors from guilds for directors, writers, producers and actors.
Despite an ending that some found unsatisfying or even baffling for its loose ends, the film has won universal acclaim and is the Coen brothers' biggest commercial success, with $60 million and climbing at the domestic box office.
Adapted from Cormac McCarthy's novel, "No Country" is anchored by three truly great performances from Josh Brolin as a wily Texan on the run with a fortune in hijacked drug money, Tommy Lee Jones as an honorable sheriff mystified by the case's brutality, and Javier Bardem as an inhuman killer trying to retrieve the cash.
The Coens' chief worry was that Bardem's character might "come across as sort of like the terminator in a bad action movie. The implacable-killer cliche, the unstoppable killer," Joel Coen said. "The thing we knew about Javier is that whatever it is he did, it would not be that."
(See below for why Bardem is a shoo-in to win supporting actor.)
Strangely (perhaps fortuitously) for the Coens, whose career has been built on oddball pictures that normally aren't the stuff Oscar dreams are made of, "No Country" is up against an even weirder picture, Paul Thomas Anderson's oil-boom epic "There Will Be Blood."
"No Country" looks almost conventional next to "There Will Be Blood." A virtual silent film for the first 15 minutes, "Blood" then hurtles through a mythic struggle between commerce and Christianity, both equally greedy and underhanded.
"Atonement" is an all-around fine film but not one people feel passionate about (the two attractive and deserving leads, Keira Knightley and James McAvoy, didn't even earn acting nominations).
Despite the supreme charm of George Clooney in the title role, the legal drama "Michael Clayton" looks to be merely along for the best-picture ride. Likewise, the pregnancy comedy "Juno" seems a just-happy-to-be-nominated entry, much like another lighter nominee, "Little Miss Sunshine," was a year ago.
Weird as it is, "There Will Be Blood" features a monumental performance from Daniel Day-Lewis, a best-actor winner for "My Left Foot" who seems certain to win again this time.
As a pioneering oil baron, Day-Lewis gives the sort of performance for which the phrase "larger than life" was invented. He's big and bad, callous and contemptuous, and riveting for every second he's on screen.
Where did all that diabolical energy come from?
"It was a fully imagined, fully understood world that Paul had already created on the page for me, therefore it was that world, in its entirety, that unleashed a curiosity that can take you, you just don't know where," Day-Lewis said.
If he did not already own an Oscar, past supporting-actor winner Clooney might have a better shot against Day-Lewis. Clooney is tremendous as an attorney rediscovering his conscience, which has gone south amid his work as a "fixer" of sordid problems.
"No Country" co-star Jones was a surprise nominee as an ex-military policeman looking into the death of his son in "In the Valley of Elah," a murder mystery set among returning Iraq veterans. A past Oscar recipient, Jones is unlikely to win again for a film generally dismissed by critics and awards watchers.
Johnny Depp is deliciously disturbed as the murderous title figure in "Sweeney Todd: The Demon Barber of Fleet Street," while Viggo Mortensen strikes a great balance between compassion and ferocity as a Russian mobster in "Eastern Promises."
But no one can touch Day-Lewis this year.
It took a lot of arm-twisting by director Sarah Polley before Julie Christie agreed to star in "Away From Her," a heartbreaking drama about a woman succumbing to Alzheimer's.
After Christie finally said yes, she went on to deliver a performance that beautifully bookends her Oscar-winning role 42 years earlier in "Darling" as a social-climber shallowly sleeping her way to the top.
In "Away From Her," Christie plays a woman with all her romantic slings and arrows behind her, who finally has arrived at a place of peace and companionship only to have it slip away as her memory fades.
The reclusive Christie understands why her performance has drawn so much attention. Everyone, Oscar voters included, loves a comeback.
"They love to see you're still alive, and not only still alive, but you're actually doing something. 'Great, magnificent, you're alive and walking. Fantastic,'" Christie said.
Oscar voters also love fresh faces, and there are two potential spoilers for best actress: Marion Cotillard, who undergoes a remarkable transformation from dauntless teen to worldly chanteuse to frail has-been as singer Edith Piaf in "La Vie En Rose"; and Ellen Page as a whipsmart pregnant teen with a bag full of smart-alecky put-downs in "Juno."
Cate Blanchett is grandly showy as Britain's monarch in "Elizabeth: The Golden Age," but it's a been-there, done-that sequel to the film that shot her to stardom. Plus she already has a supporting-actress Oscar.
Laura Linney, nominated for the sibling comic drama "The Savages," is great as usual, but with Christie, Cotillard and Page in the race, this is not her year.
Supporting actor
True evil is hard to embrace, but Javier Bardem cannot lose. His "No Country for Old Men" killer is a merciless terror out of your worst nightmares, yet Bardem imbues him with a twisted charm and tenacity that makes him irresistible and even perversely human, albeit from the depraved side of humanity.
"It's about playing somebody that is numb with any other people's feelings or even his own feelings," Bardem said. "He doesn't have any goal or any need or wish in doing what he does. He's about to bring fate to you in order for you to face it, whether you like it or not."
Turns out, everybody liked it. Bardem has dominated at earlier awards and is considered a lock to win in a category crowded with worthy performances: Casey Affleck as a jilted devotee who turns on his idol in "The Assassination of Jesse James by the Coward Robert Ford"; Hal Holbrook as a lonely widower who tries to become surrogate father to an idealistic wanderer in "Into the Wild"; past best-actor winner Philip Seymour Hoffman as a slovenly but capable CIA man in "Charlie Wilson's War"; and Tom Wilkinson as a brilliant but unstable attorney in "Michael Clayton."
In any other year, any one of them could win.
Supporting actress
Here's the category that still holds real suspense. If Blanchett had not already won an Oscar, she'd be the likely pick to take the prize for her transgender brilliance in a spot-on incarnation of Bob Dylan in "I'm Not There." But the movie itself left viewers cold, and Blanchett's win as Katharine Hepburn in "The Aviator" is fresh in Oscar voters' minds.
Child actors have fared best in the supporting-actress category, a potential plus for teenager Saoirse Ronan as a jealous girl in "Atonement" whose fib has devastating consequences for an older sister and her new lover.
On the other end is Ruby Dee, who has a fairly fleeting role as mother to a crime lord in "American Gangster" but may have sentiment on her side with her first Oscar nomination coming at age 83, after a venerable 60-year career.
"They didn't notice me till I got to be an old, old lady," Dee joked.
With "Michael Clayton" co-stars Clooney and Wilkinson facing seemingly insurmountable competition in their categories, Tilda Swinton could be the most likely performer from that movie to win for her role as an attorney who stops at nothing to achieve her ends. Next to Clooney and Wilkinson, though, Swinton's is the weakest performance of the movie's three principals.
Then there's Amy Ryan, a theater and TV veteran whose film career has taken off on the strength of her turn in "Gone Baby Gone" as the mother of all bad mothers, a woman who responds to her 4-year-old daughter's abduction with infuriating selfishness.
"Mostly, when I do a scene, I'm relieved when it's over," Ryan said. "With this, it was, 'Oh, man, I want to go again. Let's go again.' There's so much to do with her. There are so many variables to the equation."
The category is full of first-time directing contenders, with only Joel Coen having previously been nominated, for "Fargo" (he and brother Ethan used to split the producing and directing credits but lately have been sharing both).
They are up against Anderson for "There Will Be Blood," Tony Gilroy for "Michael Clayton," Jason Reitman for "Juno" and Julian Schnabel for "The Diving Bell and the Butterfly," adapted from the memoir of French Elle editor Jean-Dominique Bauby, who suffered a paralyzing stroke.
The Coens have gradually advanced from admired outsiders to exalted players on the mainstream fringes. With "No Country," they have delivered their most popular film yet, one that has found an audience at large along with the enthusiastic respect of critics.
So it seems inevitable that they will take home Hollywood's biggest filmmaking honor, which usually goes hand-in-hand with best picture.
Accepting the Directors Guild of America prize for "No Country," Joel Coen said it would find an honored place among the "various plaques and such that we've gotten over the years that we call our ego corner." On a bad day, he joked, brother Ethan goes over and shines up their trophies.
After Sunday, the Coens' ego corner should have a new centerpiece.


How do you organize you SCSS files?
Is there a class for tabs like the .even5 for button groups?
Will is be left in Foundation 6, this could be a community supported component.
Or will it be completely removed from Foundation 6.
Make sure your menu's ul has a position relative or absolute.
Then try giving it a z-index of 99.
If you could give us a link I could better inspect the styling and see if something catches my eye on what else could be causing it.
For the time being I added a 20% width to the tabs.
I will work on a more elegant solution when I come back to this section.
The final tabs will have some spacing and the highlighted tab will connect to the content.


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Nico Adomako has German and Ghanaian roots, the latter having led him to West Africa multiple times.
When asked, the DJ and co-founder of music collective Einhundert states that Berlin and Ghana aren't "comparable at all, besides the growing rate of gentrification and the influence of the outside world." Adomako is set to visit the country again in the near future, this time he hopes to play there. "It's finally infiltrating the western world, especially in cities known for their musical impact, such as London," he notes about Ghana's growing music scene.
My background is definitely part of my style and musical tastes right now. I grew up with a lot of music in my life. Mine was a mixed race household so I was always exposed to different cultures and music. My mom was into techno and hip-hop and my dad was reggae and Ghanaian music, but both are really big fans of Bob Marley who was my favorite artist. I always had the passion for music and I made cassettes for people in my class and even for the teachers at one point. I have a radio show and sometimes my mom is like, "I showed you these tunes when you were young and you didn't like them!" It's really funny.
What's special about Ghana from your perspective as a musician?
It's interesting how fast it's growing industry-wise. There's a great spirit there as well. Everyone is very welcoming and there are so many opportunities to do stuff that can't be done here anymore, I would say. The new music scene in Ghana and West Africa is right now experiencing its biggest breakthrough, various artists from there are finally getting airtime and touring. For example, Bryte, who I'm a big fan of, will play at Berghain here in Berlin for CTM festival soon.
Is there something the music scene has that's missing elsewhere, and in Berlin in particular?
Music began in Africa and this is still the spirit that is felt there; it plays such a major role in the lives of everyone. I really like how the new sound of Ghana is a mixture of Afrobeat, classical highlife music—which my dad raised me on—rap music and club sounds.
How do you feel about Berlin's eclectic nature, has it changed?
Now people actually grow up with exposure to different stuff. I would say what's changed is that the youth go to a rap concert one day and the next day they go to Berghain, and when I was young you definitely just stayed in one lane. I like to see that everyone has this energy and that they're being actually creative and less scared to enjoy or try different things.
As well as working as a DJ you also work with a lot of emerging artists that you have discovered.
I wouldn't take credit for discovering anyone. It's more about giving deserving artists a platform to showcase their work and vision, as well as trying to guide them by sharing my experiences.
With Einhundert Sound System, which is part of your music platform Einhundert, you focus your energy on local talent. Why is that important?
It's not all focused on local talents, for example we have JJAQ from Rotterdam in our Soundsystem, but we are trying to mainly give an audience to people who are based here. I think it's important to shape Berlin's music culture for the future, especially for kids who are born and raised in here, such as Low.Vision. I think it's tougher for them; the people who move here create a lot of opportunities but they also take up a lot of space from local talents.
What kind of challenges do they, and you, face?
There are a lot of setbacks involved, you're constantly faced with the challenge of how to reinvent yourself in a way that attracts a new demographic. Translating ideas to companies can be difficult when money is involved as the original vision can be easily lost sight of in favor of marketing strategies.
What is the biggest, most lasting impact Berlin has had on you?
Due to these challenges, Berlin is often frustrating as a creative, but it definitely gave me a drive to just create stuff on my own. Berlin as a melting pot exposed me to all kinds of different cultures, knowledge, and music genres. It also pushed me out of my comfort zone and helped to keep me open for all kinds of new stuff that's coming my way, such as taking the consulting agency, Bird on a Wire, that my friend Flo Hattenbach and I run together, to the next level. We're putting together another tour for Asia and North America, and launching a new event series.
And what impact do you hope to have?
I hope I can inspire someone to take a risk, follow his or her passion and find a way to get paid for it, and wait for no one!
It sounds like Berlin is, and will stay, home for you?
Berlin is definitely the place where I love to express myself and see other people doing the same. When it comes to Münster, besides my mom and brother still living there, I hardly relate to it anymore. It barely feels like home when I visit, more like a place that I know really well. Home is more like a feeling to me rather than a specific place.
Can you see yourself living anywhere else in the world?
Somewhere sunny! Maybe somewhere quiet but then I don't want to get bored. It would probably take 20 years to know where I want to go, but at the moment Berlin is my favorite city. People can follow their passions here, at least for now. This will probably change in the next five to ten years as it gets more expensive.


Ever since I was a kid I have had a fascination with all things horror. I know, I'm a weirdo but it's true. I dig what lurks in the darkness and things that go bump in the night. I grew up in Fennville, MI and my house was haunted. Now, I know that not everyone believes in ghosts or haunted houses but this home was definitely haunted. When you're able to hear kids asking for help through the vents, it's something that lets you know you're not alone. There's more to my house but I won't go into it here. I already wrote a post on it but if you would like to read up on the scary things that happened you can click here. Throughout my childhood I watched a lot of scary movies. It seemed to go hand in hand with what I experienced at my house so maybe that's why I'm such a big fan of horror. Either way though, as I grew older the games I played took a turn with the Survival Horror genre. It started with Resident Evil. I was a huge fan of trying to survive a zombie apocalypse and though the game was awesome it still didn't quench my thirst for true horror. I searched for something that would scare me and match what I felt growing up at home but nothing worked. Finally, during my senior year of high school I found a survival horror game that fit the bill. It was 1999 when Silent Hill came out. The buzz had been going around that this was Konami's answer to Capcom's Resident Evil. I bought the game in hopes that the buzz would be true. My friend Adam and I started the game and right off the bat the game was chilling and scary. What made this possible were the effects and camera shots. It was amazing how smooth the gameplay was and how the creators used different camera shots as if someone was actually watching your character walk around. The game is based on a regular guy named Harry Mason who is in search of his missing daughter in the abandoned town of Silent Hill. As he makes his way through the city he finds out that there is a connection between his daughter and the town. There is also a thick fog that fills Silent Hill and at times it appears to be snowing. Odd monsters roam the streets as well as rabid dogs. It was different from the norm and it actually scared me.
I think what scared me the most was going into the school and the hospital. The school is empty but what you hear is a different story. Children crying and laughing fill different rooms. Doors open and shut by themselves and throughout the game there is a siren that goes off. The town starts out looking spooky but once the siren goes off it morphs into something you would imagine Hell looking like. During this time there are more monters lurking and lunging at you. There are gory scenes and once you get to the hospital they get worse. Now earlier I mentioned snow but it turns out that what appears to be snow is actually ash thats falling from the sky. The town of Silent Hill has a violent and horrific past that involves a massive fire. It's pretty interesting and it made me wonder whether or not it was based on something within our history. One day I decided to search for Silent Hill as a real town and what I found was a city named Centralia. Located in Pennsylvania, Centralia was once a booming town due to its coal mining. All was good until May of 1962 when an abandoned strip mine caught fire. There are many theories as to how it started but in the end the local fire department was unable to put it out. The strip mine underneath the town of Centralia burned and caused massive damage. It was enough that the government paid to relocate the residents of this now inhabitable town but some stayed behind. There is much more to this story and it's as simple as a Google search to find the rest. This town, from what I've read, is the basis for Silent Hill. To this day the fires still burn underneath Centralia.
My search for something truly horrifying lead me to the game Silent Hill. From there I found out that it was based on a real town that suffered from a tragic fire. It's funny how something as small as searching for a horror fix could lead to something so interesting. Maybe I'll try and piece together more games or movies that are based on true events. Anyone have some insight here? I'm wondering if "The Town That Dreaded Sundown" is real? If anyone has any info on this or other interesting stories on ghost towns let me know. For now, make sure to check out the Silent Hill franchise and the video I provided below on Centralia. Happy reading folks!


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To get the best deals on Ipad Pro 6th Gen, you can head over to LazMall, which is a partnership with thousands of trusted sellers and leading brands. Besides offering the official stores of leading brands, LazMall also features a variety of independent sellers that are dedicated to providing goods and services to customers. This is because LazMall makes sure to provide every customer with a wide range of choices, including the new Apple tablet.
The iPad Air 5th Generation offers a standard 10.9-inch Liquid Retina display, along with a 12-megapixel ultra-wide front-facing camera. Its screen features 264 ppi pixel density and is also anti-reflective. It also has a faster USB-C port and a 5G modem option. Overall, the iPad Air 5th Gen is a great tablet for the price.
The iPad Air has a 3.2 GHz Apple M1 processor with eight cores and a 16-core Neural Engine. It also offers 64 or 256 GB of flash storage and a Touch ID authentication sensor. It also 홈가전 supports the Apple Pencil 2nd Gen. This stylus is sold separately and costs $129. A+ BBB ratings make cash for your Mac a snap. It can be sold to online stores that accept
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The iPad Air 5th Gen has been rumored to feature a fingerprint sensor on the back, so it's important to keep an eye out for a new release. It's the perfect tablet for travelers and those who want to enjoy music on the go. The iPad Air 5th Gen is a great upgrade from the iPad Air 2 and is the easiest to recommend within Apple's ecosystem. The tablet features a 10.9-inch Liquid Retina display and the Apple M1 chip. It is compatible with many accessories from the iPad Air 4th generation, but old iPad Air cases won't fit the new model.
The new iPad Pro models feature almost identical specs but differ by about $300. The new models have larger displays and a mini-LED backlighting, but the only difference is the price. Some accessories are cheaper for the bigger iPad Pro, but not always. The latest iPads offer a high-resolution display with HDR. For more information, visit apple.com/iPad. Alternatively, see our full review of the new iPad Pro models.
The iPad Pro is Apple's premium professional sub-lineup. It runs an optimized version of iOS called iPadOS. It was introduced in September 2015 and features a new array sensor called Face ID. This new model also supports the Magic Keyboard. The A10X Fusion processor and up to 512GB of storage make it a powerful computer. It has many advantages over its predecessors, and is a great option for anyone who needs to work on the go.
iPad Pro 12.9-inch with mini-LED display
Rumors have it that Apple is planning to release a new iPad Pro with a mini-LED display next year. While Kuo previously backed this theory, the new 12.9-inch iPad Pro could have a smaller screen with improved brightness and contrast levels. In the meantime, we'll have to wait and see. Let's take a look at some of the other features of the new iPad.
The most notable innovation of the new iPad Pro is its mini-LED display. It boasts a Liquid Retina XDR display with over 10,000 Mini LEDs. The mini LEDs create two-thousand local dimming zones. That means that it can offer better black levels and high dynamic range video playback. As a result, the iPad Pro's display will be more vivid than ever before.
iPad Pro 12.9-inch with OLED display
Several sources indicate that Apple is working on an OLED display for its new iPad Pro. The company is currently working with LG Display and Samsung to make panels for both the 12.9-inch and 11-inch models. However, it may be several years before we see a new model with an OLED display. In the meantime, the current iPad Pro models are equipped with mini-LED technology. If this is the case, the new OLED model may be a bit pricey.
According to Digitimes, Apple is working with LG Display to produce a new version of the iPad Pro with an OLED display. This new display technology allows Apple to manufacture panels of a higher brightness and longer life. It also allows it to use a variable refresh rate, which reduces the overall power consumption. It's possible that the OLED model for the iPad Pro may debut in late 2023 or early 2024.


Azospirillum brasilense is a nitrogen-fixing bacterium associated with important agricultural crops such as rice, wheat and maize. The expression of genes responsible for nitrogen fixation (nif genes) in this bacterium is dependent on the transcriptional activator NifA. This protein contains three structural domains: the N-terminal domain is responsible for the negative control by fixed nitrogen; the central domain interacts with the RNA polymerase σ54 factor and the C-terminal domain is involved in DNA binding. The central and C-terminal domains are linked by the interdomain linker (IDL). A conserved four-cysteine motif encompassing the end of the central domain and the IDL is probably involved in the oxygen-sensitivity of NifA. In the present study, we have expressed, purified and characterized an N-truncated form of A. brasilense NifA. The protein expression was carried out in Escherichia coli and the N-truncated NifA protein was purified by chromatography using an affinity metal-chelating resin followed by a heparin-bound resin. Protein homogeneity was determined by densitometric analysis. The N-truncated protein activated in vivo nifH::lacZ transcription regardless of fixed nitrogen concentration (absence or presence of 20 mM NH4Cl) but only under low oxygen levels. On the other hand, the aerobically purified N-truncated NifA protein bound to the nifB promoter, as demonstrated by an electrophoretic mobility shift assay, implying that DNA-binding activity is not strictly controlled by oxygen levels. Our data show that, while the N-truncated NifA is inactive in vivo under aerobic conditions, it still retains DNA-binding activity, suggesting that the oxidized form of NifA bound to DNA is not competent to activate transcription.
The α-proteobacterium Azospirillum brasilense is a diazotroph found in association with important agricultural crops (1). This bacterium has been recently recommended as a plant growth-promoting inoculant for maize and wheat in Brazil. A. brasilense is able to reduce dinitrogen to ammonium by the nitrogenase enzyme complex, which consists of two proteins: dinitrogenase reductase and dinitrogenase, encoded by the nifHDK genes. In order to have an active nitrogenase enzyme, other nif gene products are necessary (2). In A. brasilense, as well as in many other diazotrophs, the nif genes are transcribed by the RNA polymerase holoenzyme containing the σ54 factor, which also requires a specific transcriptional activator, i.e., the NifA protein (1). NifA binds to an upstream activator sequence, characterized by a TGTN10ACA motif, which is located about 100 bp upstream from the nif gene promoters (3). The NifA then contacts the σ54-RNA polymerase holoenzyme bound to the downstream promoter through a DNA loop and catalyzes open complex formation in an ATP-driven reaction (2). NifA activity is controlled by ammonium and oxygen in diazotrophic proteobacteria allowing nif expression when nitrogen fixation is required. In the α-proteobacterium A. brasilense, this regulation does not involve the anti-activator NifL, which controls NifA in most of the γ-proteobacteria such as Klebsiella pneumoniae and Azotobacter vinelandii (1,2).
factor, showing an AAA motif involved in ATP hydrolysis (8). The carboxy-terminal domain (C-terminal) has about 65 to 130 amino acids and contains a conserved helix-turn-helix motif responsible for DNA binding (2,4). The N-terminal and the central domains are linked by the Q interlinker (QL) sequence. The central and C-terminal domains are linked by an inter-domain region, named IDL. In A. brasilense, as well as in other members of the α-proteobacteria, at the end of the central domain and part of the IDL inter-domain there is a cysteine motif most probably involved in oxygen sensitivity (5).
In this study, we describe the expression, purification and in vitro and in vivo characterization of an N-truncated form of the A. brasilense NifA protein.
Chemicals were analytical or molecular biology grade and were purchased from Merck (Germany), J.T. Baker (Netherlands), Sigma (USA) or Invitrogen (USA). Restriction enzymes were from Fermentas/Thermo Scientific (USA) or Invitrogen. Oligonucleotides were from Invitrogen or IDT (USA). HiTrap chelating chromatographic columns and HiTrap heparin were furnished by GE Healthcare (Sweden).
The sequence encoding the central and C-terminal domains of the NifA protein was amplified using the primers Ab5'CCT (5'-CCCATATGAACGACGTGGTCTGCAC-3') and Ab3'CCT (5'-GGTGGATCCCGCGTACATGAT-3'), which respectively introduced NdeI and BamHI restriction sites (underlined). The amplified sequence corresponds to the 1038 to 2383 nucleotide according to the A. brasilense nifA sequence placed in GenBank (accession No. X60714.1). This sequence encodes the A. brasilense NifA protein lacking its first 201 amino acids, which include the N-terminal domain and part of the QL region. The amplified fragment (ΔN-nifA) was cloned into the vector pTZ57R/T (Thermo Scientific), yielding pCN57RCCT. This plasmid was then cleaved with NdeI and BamHI and the released fragment was subcloned into the pET28a vector (Novagen, USA), generating the recombinant plasmid pCNpETCCT. To construct pCNK6CCT, ΔN-nifA was released from pCNpETCCT as an XbaI/HindIII fragment and cloned into pDK6 (9). The amplified fragment was sequenced to confirm gene integrity.
The recombinant plasmid pCNpETCCT was introduced into Escherichia coli strain BL21(DE3) (10), and protein induction was carried out using 0.5 mM IPTG for 16 h at 18°C in 300 mL Luria Bertani (LB) medium. After induction, cells were harvested by centrifugation at 4700 g for 10 min at 4°C, and then resuspended in 1 mL sonication buffer (0.5 mM NaCl, 50 mM Tris-HCl, pH 8.0, 10% glycerol, 1 mM DTT) per gram of wet cells in the presence of 1 mM PMSF. The cells were lysed by sonication in 10 cycles of 20 s, and the crude extract (102 mg total protein) was centrifuged at 12,000 g for 15 min at 4°C to remove cell debris and insoluble proteins.
Protein purification was performed using a 1 mL HiTrap Ni2+ chelating column equilibrated with Tp buffer (50 mM Tris-HCl, pH 8.0, 500 mM NaCl, 5% glycerol, 0.1 mM DTT) containing 50 mM imidazole. After loading the soluble protein extract (48 mg protein), the column was washed with 2 volumes of Tp with 50 mM imidazole and 2 volumes of Tp with 100 mM imidazole. The protein was eluted by a stepwise increase in imidazole concentration (200, 250, 300, 400, 1000 mM) in Tp buffer with 1 column volume of each concentration. Fractions of 1 mL were collected and analyzed by SDS-PAGE (11). The ΔN-NifA protein was eluted at 250 and 300 mM imidazole. These fractions were pooled and diluted with 1 volume of 50 mM Tris-HCl, pH 8.0, and then loaded onto a 1 mL HiTrap heparin column previously equilibrated with Tp3 buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl). After washing the column with 3 volumes of Tp3, protein was eluted by a stepwise increase in NaCl concentration (500, 600, 700, 800, 900, 1000, 1500 mM) in 50 mM Tris-HCl, pH 7.5, with 1 column volume of the first two and the last concentrations and a 1/2 column volume of the other concentrations. Protein eluted with 700 mM NaCl was used for the DNA-binding electrophoretic mobility shift assay (EMSA).
Protein was quantitated by the method of Bradford (12) using bovine serum albumin as standard. The protein homogeneity was determined by densitometric analysis of the protein bands on SDS-PAGE (11) stained with Coomassie blue R-250.
A 345-bp DNA fragment corresponding to region 2510 to 2855 of the Herbaspirillum seropedicae nifB promoter (13) was amplified using primers labeled with fluorescent FAM and VIC. The reaction contained 0.1 µM labeled-DNA (H. seropedicae nifB promoter region), DNA-binding buffer (10 mM Tris-acetate, pH 8.0, 8 mM MgCl2, 10 mM potassium acetate, 1 mM DTT, 3.5% (w/v) PEG 8000) and increasing concentrations of the purified protein in a final volume of 15 µL. The reactions were incubated for 10 min at room temperature. After incubation, 5 µL 15% glycerol containing 0.01% bromophenol blue was added and the reaction was loaded onto a 4% non-denaturing polyacrylamide gel in Tris-glycine buffer (25 mM Tris-base, 192 mM glycine) with 8 mM MgCl2. Electrophoresis was performed at 60 V for 4 h at 4°C. The gel was visualized on a transilluminator and registered with a video-image- coupled system (UVP).
E. coli JM109(DE3) (10) cells carrying plasmid pRT22 (K. pneumoniae nifH::lacZ) (14) were transformed with plasmid expressing the A. brasilense ΔN-NifA protein. Cells were grown overnight in nitrogen-free Davis and Mingioli (NFDM) medium (15) supplemented with 5% LB medium, 20 mM NH4Cl and antibiotics at 30°C. The cultures were then diluted to absorbance at 600 nm ≈0.2 in NFDM containing 0.5 mM IPTG and in the presence or absence of 20 mM NH4Cl. Cultures were incubated at 30°C for 6-7 h under anaerobiosis, with oxygen replaced with nitrogen or argon. β-galactosidase activity was determined using ONPG as described (16).
One major difficulty for in vitro analysis of the NifA protein is to obtain an adequate amount of purified protein since the protein is usually expressed at very low concentration under physiological conditions. Moreover, NifA tends to be insoluble when overexpressed, which also contributes to the difficulty of its in vitro characterization. The overexpression of the full-length A. brasilense NifA in E. coli also yielded an insoluble protein regardless of the different conditions used for expression and cell lysis (data not shown). Thus, in the present study, we removed the N-terminal domain of the NifA protein in order to improve solubility of the overexpressed protein. The N-terminal domain is not strictly required for the transcriptional activation, although it is involved in ammonium regulation (6).
The N-truncated NifA protein (ΔN-NifA) was first assayed for in vivo transcriptional activation activity in E. coli JM109(DE3) carrying plasmids pRT22 (nifH::lacZ) and pCNK6CCT (ΔN-NifA expressed from the tac promoter). These assays were carried out in the presence or absence of 20 mM NH4Cl, and under air (21% O2) or anaerobiosis (Table 1). β-galactosidase activity was only observed in cells carrying both plasmids, in the absence of O2, indicating that the ΔN-NifA protein is able to activate nifH transcription under these conditions. Regulation by ammonium ion was not observed under these conditions, reinforcing the evidence that the N-terminal domain is involved in ammonium control but not required for transcriptional activation (6). On the other hand, regulation by O2 was retained by the ΔN-NifA, most probably because the cysteine motif is still present in this form.
The purification of ΔN-NifA from A. brasilense fused to a His-tag sequence was obtained using plasmid pCNpETCCT and expression in E. coli BL21(DE3). By performing protein induction at 18°C, about 50% of the expressed protein remained in the soluble fraction of the crude extract, allowing its purification. Two chromatography steps were used: a metal-binding affinity chromatography, using a HiTrap Ni2+ chelating column, followed by HiTrap heparin chromatography (Figure 1). This chromatographic procedure yielded 80% pure protein as determined by densitometric analysis of Coomassie blue-stained SDS-PAGE.
The purified ΔN-NifA was assayed for in vitro DNA-binding activity to the H. seropedicae nifB gene promoter region using an electrophoretic mobility shift assay (Figure 2). The NifB protein is involved in the synthesis of the nitrogenase FeMo co-factor, and the nifB promoter contains a fully characterized NifA binding site (13). In this assay, the fluorescent labeled-DNA containing the nifB promoter showed a slower electrophoretic migration in the presence of increasing amounts of the purified A. brasilense ΔN-NifA, indicating that the protein was able to bind to the nif promoter DNA. At a protein/DNA ratio of about 12 (1.2 µM ΔN-NifA), a DNA-protein complex was clearly observed, and in the presence of 3.2 µM ΔN-NifA, the free DNA band was not observed. This DNA-protein complex was also observed even in the presence of ten times mass excess of calf thymus DNA, although some of the DNA-protein complex was disrupted at higher concentrations of calf thymus DNA (data not shown). These results indicate that the purified ΔN-NifA protein from A. brasilense was able to bind to the promoter region of nifB from H. seropedicae in the presence of oxygen, implying that the DNA-binding activity of NifA is not strictly oxygen sensitive, although higher activity and/or specificity may depend on the oxygen level. Passaglia et al. (17) have previously shown that A. brasilense NifA was capable of binding nifH in vitro under aerobic conditions. In contrast to our study, these investigators used a crude extract of E. coli expressing a full-length A. brasilense NifA, and they also concluded that the DNA-binding activity was not oxygen sensitive. Nevertheless, to the best of our knowledge, no other report has shown in vitro DNA-binding activity using purified A. brasilense NifA.
Models for the regulation of NifA activity of A. brasilense and H. seropedicae in response to the levels of ammonium and oxygen have been proposed (6,7,18,19). According to these models, the N-terminal domain folds over the catalytic central domain inhibiting transcription activation under high ammonium conditions. In the presence of low ammonium concentrations, a PII protein (GlnB for A. brasilense or GlnK for H. seropedicae) interacts with the N-terminal domain, relieving the inhibition. Our results agree with these models since we did not observe ammonium regulation or dependence on the co-expression of the A. brasilense GlnB protein. Regulation by oxygen, in turn, has been proposed to rely on a probable Fe-S cluster located in the central domain and IDL linker (5). Monteiro et al. (7), using an N-truncated form of H. seropedicae NifA, also showed that an aerobically purified NifA retained DNA-binding activity. Moreover, Oliveira et al. (18) produced mutants for each of the four cysteine residues of the putative Fe-S cluster binding motif in H. seropedicae NifA, and observed that the mutant proteins were still able to bind DNA although the ability to activate transcription was completely abolished. Oxidation of the Fe-S cluster may trigger conformational changes affecting nucleotide binding and/or hydrolysis necessary for transcriptional activation, as proposed (18,20). Therefore, under aerobic conditions, the oxidized Fe-S cluster would lead to a transcriptionally inactive form of NifA possibly compromising the catalytic activity of the central domain. It is possible that the affinity of the oxidized NifA for nif promoters is also decreased, but this possibility has not been experimentally tested. Our data show that while the N-truncated NifA is inactive in vivo under aerobic conditions, it still retains DNA-binding activity, suggesting that the oxidized form of NifA bound to DNA is not competent to activate transcription. A similar result was also reported for the H. seropedicae NifA, thus raising the possibility that a common mechanism of oxygen-dependent NifA activity control operates in both A. brasilense and H. seropedicae.
Figure 1. SDS-PAGE analysis of the Azospirillum brasilense ΔN-NifA protein expressed in Escherichia coli BL21(DE3) and purified as indicated in Material and Methods. Panel A, Crude extract (lane 1) and protein fractions eluted with 250 and 300 mM imidazole (lanes 2 and 3, respectively) from HiTrap Ni2+ chelating columns. Panel B, Protein fractions eluted with 700 and 800 mM NaCl (lanes 1 and 2, respectively) from HiTrap heparin columns. Arrows indicate the ΔN-NifA. Molecular masses are reported in kDa. Proteins were stained with Coomassie blue. Gels were 10% polyacrylamide in SDS-PAGE.
Figure 2. Electrophoretic mobility shift assay using the purified ΔN-NifA protein and Herbaspirillum seropedicae nifB promoter region. The procedure was performed as described in Material and Methods. The nifB promoter region was labeled with the fluorophores FAM and VIC and incubated with increasing concentrations of purified His-N-truncated NifA (lane 1, absence of ΔN-NifA; lanes 2 to 7, 0.16, 0.4, 0.8, 1.2, 1.6, 3.2 µM ΔN-NifA, respectively). Arrows indicate unbound DNA and the DNA-protein complex.
Table 1. Promoter activity of Klebsiella pneumoniae nifH::lacZ fusion in Escherichia coli JM109(DE3) cells.
We are grateful to Roseli Prado, Julieta Pie, Marilza Lamour, Valter Baura, and Patrícia Sotomaior for technical assistance. We are also grateful to Dr. M. Geoffrey Yates (UK) for reading the manuscript. Research supported by INCT-Fixação Biológica de Nitrogênio, CNPq, CAPES, and Institutos do Milênio.
Received September 6, 2011. Accepted January 9, 2012. Available online January 27, 2012. Published February 17, 2012.


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Q: How to I check what the php environment variables are currently set to on AWS EB? How can I check what the PHP environment variables below are currently set to on my AWS EB (Elastic Beanstalk) instance?
upload_max_filesize =
post_max_size =
memory_limit =
max_execution_time =
max_input_time =

A: You can login to the instance with the command eb ssh. This will temporary open port 22 and login to the instance via ssh.
http://docs.aws.amazon.com/elasticbeanstalk/latest/dg/eb3-ssh.html
Now you can check or update your php.ini file.
You can also create a page or run phpinfo() from command line:
php -r 'echo phpinfo();'

redpajama_set_name: RedPajamaStackExchange