text:
- Actor-Shared Background: In "Connection Lost" we're told that Matt's mother is from Karachi, a background borrowed from his actor Geoffrey Arend.
- California Doubling: Naturally there's quite a bit of it used in the series—the McCords' horse farm is said to be in Virginia but is filmed on an estate in Long Island that also plays the role of a public park in "Ghost Detainee", to say nothing of international locations—but the scenes at the UN in season 2 were filmed at the actual UN General Assembly building in New York City, averting the trope.
- The Cast Showoff: In "The Show Must Go On", Nadine, Blake, and Daisy sing Billy Joel's "The Longest Time" (with the lyrics rewritten by Matt to be about a trade deal) at a conference. Nadine (Broadway leading lady Bebe Neuwirth) leads with a very nice alto croon, and Daisy (Patina Miller) is no slouch either, her actress having won a Tony award for Pippin shortly before joining the show.
- The Danza: Played with. Elizabeth McCord is played by Téa Leoni — whose actual first name is, in fact, Elizabeth.
- Dmitri Petrov is played by Chris Petrovski.
- Fake American: eljko Ivanek (Russell Jackson) is originally from Slovenia, though he's been an American citizen for years.
- Fake Nationality:
- Usman Ally (Iranian Foreign Minister Zahed Javani) is from Swaziland.
- Ironically, Ukrainian President Mikhail Bozek is played by a Russian-born actor, Lev Gorn.
- Fake Russian: Olek Krupa (Polish) and Yorgo Constantine (Greek-American)respectively play Russian President Pavel Ostrov and Foreign Minister Anton Gorev. Chris Petrovski (Russian Army Captain Dmitri Petrov) is originally from Macedonia. Angela Gots (Maria Ostrova) is a downplayed example: she was born and raised in New York but her parents are originally from St. Petersburg. Another New Yorker, Nicholas Guest, plays Ostrova's successor President Salnikov.
- Life Imitates Art: "Catch and Release" ends with "Jihadi Judd", an American who joined ISIS, being killed in a drone strike on orders of President Dalton. His model, British-born ISIS member Mohammed Emwazi, a.k.a. "Jihadi John", was killed in a joint US-British drone attack four days after the episode aired.
- Queer Character, Queer Actor: Kat Sandoval and actress Sara Ramirez are both bisexual and queer - in fact, Kat's coming out conversation was pretty much just Sara explaining all the details of her own personal intersectional identity.
- Romance on the Set: According to People magazine, Téa Leoni and Tim Daly's chemistry as Liz and Henry McCord wasn't all acting: they apparently started dating in December 2014, midway through season one. They're still together to this day.
Trivia / Madam Secretary
Go To


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After School Game Changer Practitioner Training
The After School Game Changer Practitioner Training will run from the 8th to the 11th of July from 08h30 to 16h30 daily, RSVP to firstname.lastname@example.com by the 21st of June as space is limited.
The After School Game Changer Practitioner Training will run from the 8th to the 11th of July from 08h30 to 16h30 daily, RSVP to firstname.lastname@example.com by the 21st of June as space is limited.
There are often a number of difficulties, complexities and curiosities that organisational leaders are faced with and do not get the space to collectively discuss.
Join fellow Directors in a two-hour conversation to craft a platform through which NPO Directors in the After School space can learn, share, network, and explore a range of common issues as determined by you.
Please note that this event is by invite only, contact firstname.lastname@example.com to RSVP.
We look forward to your engagement!
There are often a number of unique complexities and curiosities that organisational leaders face on a daily basis and do not get the chance to talk about in like-minded spaces.
The Learning Trust will be hosting a Director's Circle for Directors of our grantee partners and Alumni in the Western Cape on Friday the 31st of May.
The sessions are by invite only, for any queries contact: email@example.com.
See you there!
Join us at the second OASIS CoP in this year's season where we finalise the plans towards our Pilot Project Implementation, the OASIS School of Dialogue and revisions to our Mobile App. See below the agenda for the day:
Review of OCT Action Points
OASIS School of Dialogue
OASIS Mobile App
OASIS Pilot Project
Funding Proposal
MoU - The Learning Trust
Response from The Learning Trust Executive Director, Sibongile Khumalo
RSVP to Mark Lamani at firstname.lastname@example.com
Looking forward to our engagement!
Do you want to better understand why learners attend your After School Programme and how you can reach those learners who don't?
Join us at the upcoming Community of Practice event to share and gain valuable insights, information and networks as we address the questions:
What attracts learner's to after school programmes?
How are after school programmes maintaining participation over time?
What are the latest South African research findings?
RSVP here as space is limited.
For enquiries contact: email@example.com
After School Programmes are highly underestimated and undervalued in terms of the developmental benefits they hold for child and youth participants as well as young adults employed in the sector.
More than just a place of safety, it is often a hub of activity defined by human connection. Whether it's an academic tutoring programme, reading club, sports game or drama & dance class, young people participating in extended learning activities get to connect with each other. In such spaces they are given the opportunity to enjoy a participatory activity that is filled with fun, an opportunity to express themselves, to be acknowledged, and to acknowledge and help others while learning a new skill.
In addition, these young people connect with adult after school practitioners (tutors, coaches, etc.) who take the time to acknowledge, nurture and SEE them as young people developing into greater versions of themselves.
Yet, do we understand the real impact of this work on the behaviour of a young person? And how does our valuable work in the after school space contribute to constructive behavioural change for young people living in a destructive context as a result of social and economic brutality.
Our one day seminar in 2019 seeks to help our grantees explore the above-mentioned theme. We aim to start with a morning session that will be a basic introduction to the science of Behavioural Change and an afternoon session where they get to unpack how this knowledge applies to grantees' work and how it could be used to better monitor, evaluate and articulate their value add in their programmes.
We will be having one seminar per province, as per details below…
12 March, 10h00 - 15h00. Ikamva Youth: 47 Westminster Rd, Salt River, Cape Town, 7925. (Western Cape)
14 March, 10h00 - 15h00. Ikamva Youth: 49 Boshoff Rd, President Park AH, Midrand, 1685. (Gauteng)
28 March, 10h00 - 15h00. Assumption Development Centre: Nceme St. Makhanda, 6139. (Eastern Cape)
To secure your spot at one of these seminars, please complete the RSVP form here.
Looking forward to see you there!
The Western Cape Education Department announced their grade 12 results in early January, revealing a slight drop in the overall pass rate, a higher number of bachelor passes, a higher failure rate, as well as a lower drop-out of grade 10 to 12 learners from the system. We know that many matriculants continue to fail or do badly in their final year of school, which further widens the opportunity gap for young people.
Our next CoP will focus on making the link between the opportunity gap with After School programmes (or extended learning/support for in-school learners). Our speakers will talk us through their personal experience working with learners and what ASPs can do to address this gap. We will explore the link between the quality of learners coming out of the schooling system and whether they are well prepared for post-school and what ASPs can do to improve on this preparation.
Join us on Wednesday 27 February, 9am - 1pm
Isivivana Centre
8 Mzala Street (Corner of Mzala & Julius Tsholo Streets)
Khayelitsha, Cape Town
To book your place: Click here
Enquiries: firstname.lastname@example.com
We look forward to welcoming you!
The Learning Trust and the Centre for Community Schools are hosting a Stakeholder Breakfast under the banner of the OASIS (Organising After School and In School) Community of Practice in Port Elizabeth on the 28th of November 2018, 10h00 – 12h00, at Nelson Mandela University, Missionvale Campus.
For more details, please contact firstname.lastname@example.com.
The Learning Trust, in partnership with the After School Game Changer, invites you to join our fourth and final Community of Practice (CoP) meeting for 2018.
In our sector, we work with volunteers from a wide range of backgrounds. With this event, we explore strategies for attracting, up-skilling and retaining volunteers for maximum impact. Join us as we share insights on how to make volunteerism an enriching experience for all.
Venue: Philippi Village, Cwangco Crescent, Philippi, 7781, Cape Town
To book your place: Click here
Enquiries: email@example.com

subset: commoncrawl


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James was amazing from start to finish. He explained the process to me in detail. And handles expectations well considering it was a down market. I highly recommend James to my friends and family, he kept me in the loop with everything going on in the deal. His communication and negotiation skills were great got us a good price for the property. ... Read more
James Wong is a absolutely outstanding experience real estate agent. Once you call him to arrange an appointment, he always show up on time. I bought the house back in year 2014, when the time he deal with seller, he came up with his absolutely responsibly to get the best price between seller and buyer. I also referral to my friends and my relatives to him, and I highly recommended him, James Wong personal real estate agent!... Read more
James is very nice and professional agent, he always pay attention to find more resource to offer to us, and helped us a lot to purchased the apartment. We highly recommend him :) ... Read more
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If anyone are trying to choose to sell or buy your property, definitely James Wang he is the one. He is very professional, very personable and knowledgeable guy. I found working with him a very comfortable experience, and profitable. He works non-stop until he gets the price that I want. He is an excellent communicator.... Read more


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Original owner, and meticulously maintained condo, located in a secure and gated community!... Read More
Owner financing available! great condition condo with 2 bedrooms and 2 full baths. kitchen... Read More
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A 2br/2ba split floor plan condo that is very clean with neutral paint. the greatroom has vaulted... Read More
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|Average Price
|2 Bed Condos For Sale in Town Creek (8)
|$ 98,350
$ 98,350
$ 88
Currently there are and 8 condos and apartments for sale in Town Creek
Town Creek is a subdivision within the city of Greensboro, Georgia. There are currently 8 Homes for Sale within Town Creek, with asking prices ranging from $82,500 to $125,000. The average list price per square foot of the available inventory in Town Creek is $88, which is below the Greensboro average of $283. The Average Sales price of all sold real estate in this subdivision for the past year is $71,250. Recent comparable homes have been selling for 93.62% of their asking price. Waterfront homes represent 0.00% of the available Town Creek inventory. The population of Town Creek, according to the 2010 Census, is 3,359. The total number of households in the zip code of Town Creek is 4,613, with an average household size for of 2.36 people. The total number of renter households within the zip code of is 1,185, which means that 74.31% of households in this zipcode are owner occupant households.
Disclaimer: Listing broker has attempted to offer accurate data, but buyers/lessees are advised to confirm all information IDX information is provided exclusively for consumers' personal, non-commercial use that it may not be used for any purpose other than to identify prospective properties consumers may be interested in purchasing, and that data is deemed reliable but is not guaranteed accurate by the MLS. The MLS may, at its discretion, require use of other disclaimers as necessary to protect Participants and/or the MLS from liability.
This data up-to-date as of [06/16/2019]


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Good clear signature on one side of slip of paper, an irregular rectangle roughly 1.5 x 12.5 cm. In fair condition, aged and worn, with traces of mount on reverse. Reads: 'Your's [sic] most affectionately | James Hogg'.
1p, 4to. Bifolium. Addressed on reverse of second leaf, with two postmarks (one from Egham) to 'Mr Carpenter | Bookseller | Old Bond Street | London'. Aged and worn, with Carpenter's spike-hole through one word of text. Reads: 'Miss Robinson will be obliged to Mr Carpenter, to send her the following works of her Mother's, uniformly bound – Together with the account of them: - Miss R must have them before Christmas day. | Viz. - | "Herbert De Sevrac" | "Vancenza" | "Angelina"'.
3pp, 12mo. Bifolium. In fair condition, aged and worn. Folded once. Wretched handwriting, resulting in the following tentative transcription. He begins by saying that he is sorry she has been ill, adding, 'I have no advantage over you in that matter. The [?] and [?] got hold upon me.' Regarding the celebrated circulating library, he writes that 'Mudie's little game is 'not to [buy?] another little [me?], knowing that he can weary out the public. | I have therefore to circulate my own copy among students, but it is out at present.
Five items in good condition, all lightly aged. ONE: ACS. Signed 'Kathy'. 1 March 1990. Clearly a covering note on sending the other material. Simply reads: 'Good to speak – | love | Kathy'. TWO: Duplicated copy of typescript of speech by 'Fr. John Gilling'. Headed: 'REQUIEM MASS: St. Mary the Virgin, Bourne Street. 7th February 1990. | JONATHAN GRIFFIN'. 2pp, 8vo.
1p, 4to. In good condition, lightly-aged. Folded three times. Sixteen-line poem, neatly typed (with one error overtyped an autograph proof mark separating two words) titled 'For Andrew Young. 1885-1971.' Signed at bottom 'Richard Church.' A charming poem, which was published in the Cornhill Magazine in 1971.
Three items (letter, photographic card, and pamphlet), all in good condition. ONE: ALS. 31 December 1954. 1p, 8vo. Signed '(Mrs.) Rita Spurr.' and addressed to 'Dear Mr. Christopher Fry'. She begins by thanking him 'for the very great pleasure & stimulus' which she derived 'during the year that is passed' from his play 'The Dark is Light Enough'.
Both items in good condition, lightly aged, as is an enclosure to the second letter, a photocopy of Lowbury's pamphlet 'A Letter from Hampstead'. ONE: 24 September 1969. 2pp, 12mo. In envelope addressed to Fry at his East Dean home The Toft. He is grateful for Fry's letter praising his collection of poems 'Figures of Light'.
2pp, landscape 12mo. In good condition, lightly aged. Begins: 'Dear Christopher Fry, | I am sorry to contradict you, but I showed my co-editor, the poet Rodney Pybus, your poem "For Edmund Blunden", and we both [last word underlined] feel it should be published, and that we would like to publish it. Please.' In the hope that Fry will agree, he asks him to 'sign and return an acceptance form and send us the biog. note you would like to print alongside the poem'. He continues: 'I do hope you'll agree.
1p, 4to. On grey paper. In good condition, lightly aged. Folded twice. With 'C. B.' date stamp at head. Addressed to 'Dear Mr. Fry'. He thanks him for his letter and 'very kind enquiry'. He is much better, 'but still not yet really well again'. His indisposition would not prevent him from sending Fry 'a poem for "Springboard" if anything conceivably suitable for it were available. But alas, this is not so; and I am sorry indeed not to be able to do what you so kindly suggest.' Valediction in autograph: 'With all good wishes to the Magazine | Yours sincerely | Walter de la Mare'.
2pp, landscape 12mo. In good condition, lightly aged. With envelope addressed by Dickinson to Fry at his East Dean address. Thirty-eight lines of closely-written text. An affectionate letter, which begins with Dickinson exclaiming, following a seasonal phone call from Fry, 'There's no-one anywhere could have given me more pleasure, yes truly so. Bless you!' He describes the illness which made the 'making of the prog […] a bit fraught'. He reports that 'The recording was strange, Lawrence had never done an unscripted interview (nor had I) nor had the producer!
Both 2pp, 12mo. In good condition, lightly aged. Written in Whistler's calligraphic hand. ONE: 13 December 1961. He will be 'so pleased' if Fry will accept a copy of his poems, 'which I like to send you because of my admiration for your work, and sense of sympathy with what you express, or create'. TWO: 21 August 1963. 33 lines of text, written following the death of his friend Christopher Hassall.
1p, landscape 12mo. In good condition, folded twice. With envelope addressed to Fry at his East Dean address. Fry has written at the head of the cover of the envelope: 'From Norman Nicholson'. After thanking Fry for his 'good wishes at Christmas', Nicholson continues: 'I, too, am sorry that we have not met, which – unless you are ever in Cumberland – seems less likely than ever, as I am much less mobile since the death of my wife.' He presumes Fry has learnt of his bereavement from Edward Lowbury, 'for I know that you were a friend of his father-in-law's'.
Six items: four from Causley (one ALS, one TLS, two ACsS) and two from Hanke (ALsS). The collection in good condition. ONE: ACS from Causley. Signed 'Charles Causley'. Launceston; 31 August 1979. He is delighted that Fry 'thought of using the poem in your reading, & hope you went ahead'. He has been 'almost 2 months in Y'slavia & Czechoslovakia'. He is 'always 5 years behind with commitments & the WC Writers' beanos almost always coincide with fearful attack of conscience. The next three items signed 'Charles'. TWO: TLS from Causley. On his Launceston letterhead; 5 August 1986.
Valerie Eliot's six letters are intimate and energetic, two of them casting an entertaining sidelight on Sir Colin St John Wilson's efforts to acquire Jacob Epstein's bust of 'Tom' for the new British Library, with a case of 'sabotage' as Wilson takes her on a tour of its new premises at Euston. The collection is in good condition. All of Valerie Eliot's letters are addressed to 'Dearest Christopher', the last in autograph. ONE: 1 September 1994. 1p, 12mo.
The relationship between Arthur Joseph Munby (1828-1910) and his wife Hannah Cullwick (1833-1909) - well described as 'one of the strangest love stories of the nineteenth century' - continues to arouse great interest. The news that - unbeknown to his own family - a well-connected Cambridge-educated barrister should have been married for almost forty years to a maidservant, was greeted with astonishment on Munby's death, the interest only heightened by the fact that Mrs Munby had refused throughout her marriage to become 'a lady'.
1p, 8vo. In good condition, lightly aged and creased. Folded once. 'I was hoping you would make a suggestion about price. I had at first thought of asking £5 for each copy, but if you would throw in that copy of my father's book (my own is in pieces) I suggest £12 for the three. Of course I want you to make your profit.' He ends by asking if Hancox would like the books signed.
Both items in fair condition, foxed and lightly aged. No indication has been discovered that the poem was published. ONE: Autograph Letter in the third person to 'Master W. A. Wilson'. 1p, 12mo. Reads: 'Sir Noël Paton has very great pleasure in responding to Master W. A. Wilson's friendly and well-written request for his Autograph. He desires to note however, that he is a Scotsman, not an "Englishman!"' Paton's compliment was perhaps backhanded, as the recipient 'Master W. A.
3pp, 12mo. Bifolium. In good condition. Folded twice. She presumes that the recipient 'may like to have my report at once for the prizes', and asks for it to be sent back in order to type it up. 'The Children's verse is', she finds, 'very even in quality which has made it so hard to judge as I feel that age must fairly be considered as between say 10 & 15.' She has decided on '10 years old as first place', despite the fact that 'the verses were not as poetical as the poem on Eclipses or the Windy Day'. Several of the children, she thinks, 'may do so much better.
Three excellent letters relating to the reclusive Antrim poet 'Moira O'Neill', mother of the novelist Molly Keane ('M. J. Farrell'): an ALS from the poet herself, explaining her reticence; an ALS from the poet John Stevenson of Coolavin ('Pat M'Carty') regarding the poet and her background, and reporting information received from the antiquary Francis Joseph Biggers and 'Rev. H D Murphy of St. George's'; and a TLS from H. C. Montgomery of Belfast, telling what he knows of the poet and her family. From the papers of Rev.
1p, 4to. In fair condition, lightly aged and worn. Folded four times. The letter refers to Robinson's 1938 memoir 'Three Houses', which tells the story of his Kinsale childhood. Begins: 'Dear Robert: | How nice to hear from you and to hear you liked "Three Houses" - I think a nice, gentle, kind book.' He remembers Wilson's mother – 'aren't all nice mothers alike?' - as well as his 'aunt in the north who likes you but was a little shocked by some poems you wrote'. He saw 'John [McGreevey?] a few weeks ago in London, and Geoffrey two summers ago'.
2pp, 12mo. On pink paper. In fair condition, lightly aged and worn. Folded twice. A letter of condolence beginning: 'Dear Robert. | I have just seen the miserable news of the death of your wife. I had heard she was ill but had no idea things were so bad.' After asking him to accept his sympathies, he continues: 'I well remember the last time I saw you both together at my studio in Dublin & I can hardly realise yet that she has gone.' He has been 'out of Dublin so long now', and so seldom sees people that he doesn't often 'hear news of folk'.
Pepys's house at Brampton is the subject of an article by Chris Partridge in the Observer, 30 May 2004, which states that 'The first earl, Edward Montagu, was Pepys's cousin and patron, giving him the political clout to further his career in the Navy Office. In 1927 the then earl gave the Pepys House Trust a 100-year lease at a peppercorn rent, and it has been open to the public ever since. All three items in good condition, lightly aged. Drinkwater's second letter and Chappell's draft reply pinned to one another. ONE: Drinkwater to Chappell, 31 May 1933. TLS. 2pp, 4to.
3pp, 12mo. Bifolium. In fair condition, lightly aged and a little grubby. Folded once. Twenty-seven lines of text. Written in a far more ragged style than Bridges's usual calligraphic hand, perhaps suggesting undermined health. Wilson is 'at liberty to quote the verses of mine that you ask for – I take it as a complement [sic]' He thanks him for 'the kind gift of your book, wh I have not had time to read'. Noting that Wilson is 'interested in sacred wells', he states that 'our "Chilswell" is probably not a well.
On one side of square of paper cut from a 12mo leaf. In fair condition, aged and lightly creased. Folded twice. Reads: 'For Coals and the School | With Lord Tennyson's | Compliments and best Wishes | May 1 / 85'. Presumably 'the School' is the Blue Coat School at Aldworth, and it seems that Tennyson was sending a cheque for the purchase of 'Coals', possibly as part of a tradition.
2pp, 12mo. In good condition, lightly aged and worn, with thin strip of paper from mount adhering to reverse of leaf. Folded three times. The recipient is not named, but the letter is annotated in pencil with the words 'From Bath – in the Spring of 1805 | To a lady of the name of Riddell' (explaining a pun on the word 'Riddle' in the letter). (Another candidate for recipient is the 'Miss Moore' referred to in the letter, although this does not accord with the salutation.) Apparently written in the final months of Currie's life.
The typed letter (2pp) and the four typescript poems (5pp) total 7pp, foolscap 8vo. In good condition, lightly aged. In good condition, lightly aged. Folded twice. LETTER: TLS to 'Dear Lawrence'. (The recipient Lawrence Wilson is less well-known as a poet than his brother Robert (or Robin) Noble Denison Wilson (1899-1953), known as R. N. D. Wilson.) 2pp, 8vo. A long letter, with seventy-three lines of typewritten text by William Montgomerie, with a long Autograph Note Signed by his wife 'Norah' at the foot and up one margin of the last leaf.
2pp, 12mo. On bifolium. In fair condition, aged and a little grubby. Folded once. Twenty-seven closely-written lines. The letter is addressed to the author James Alexander Steuart (1861-1932), and refers to his recent novel 'Wine on the Lees' (1899). There is also a reference to 'the Whitefriars men', i.e. the staff of the St James's Gazette, which was published in Dorset Street, Whitefriars.
[12]pp, landscape folio. Sewn with red thread. The words 'The Big Grey Man' on cover and title-page in red ink, rest of pamphlet in black ink. In fair condition, lightly aged and worn. A 48-line poem, in eight six-line stanzas, spread over three rectos, signed at end 'Richard Rowley.' Inscribed on title-page: 'Mrs. Brown. | With best wishes for Christmas | from | Mr. & Mrs. R. V. Williams. | 1942.' Six bucolic illustrations reproduced from woodcuts by Annesley, including one on cover. Colophon on recto of last leaf, with printers' slug on otherwise-blank reverse.
Proctor was a much loved individual in literary circles, from the days of the London Magazine to the mid-Victorian period, in which he was the dedicatee of both Thackeray's 'Vanity Fair' and Wilkie Collins's 'Woman in White'. His reputation as a poet was international: he was thought highly of by Pushkin. 1p, 12mo. In good condition, lightly aged, with thin strip of paper from mount adhering to blank reverse. The letter reads: 'Dear [Harris?] | Pray tell me where [?] I can have some conversation with you about some patients of the name of [Lealer? Lester?], whom you know.
On 17 x 12 cm leaf of thickish paper, removed from 'The Meredith Birthday Book'. In good condition, lightly aged and worn. Scratchy signature, underlined. In one of three printed boxes of red rules on one side of the leaf, the other side bearing quotations from George Meredith for the days 25 to 27 April.
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A Few Items Left
Today marks the beginning of the last week of the Kansas Legislative Session. There are still three major topics to finish up: fixing the school finance bill "error," turning back the Senate's irresponsible tax cut bill, and passing the state's budget for the next fiscal year. Here's where we are on all of these topics.Read more
ACTION - Email Legislators to Fix the Education Funding Bill the Right Way
Ask the Kansas Legislature to do right by schools and fix the education bill the right way! Do it today! They go back into session tomorrow!
By now you've heard about SB 423, the education finance bill that passed two weeks ago with an error resulting in most Kansas school districts getting less money than was intended by the bill. In total, this error reduces the amount for schools by $80 million. Some large districts, like SMSD, are slated to lose as much as $2 million, an amount described as catastrophic to schools.Read more
An Irresponsible Tax Bill
This week we present a guest column by Heidi Holliday, Executive Director of the Kansas Center for Economic Growth. KCEG has been at the forefront of demystifying tax policy and promoting responsible fiscal solutions for Kansas, and has been a partner for MainStream in economic topics. At issue in this column, the bill passed by the Senate to refuse almost $500 million in taxes owed to Kansas due to the Federal tax cuts. Quite simply, Kansas cannot afford to turn this money away. And who would be receiving this "windfall?" Not the poorest of Kansans. Not those who would most benefit from infrastructure jobs, a lowered sales tax on food, fully funded public education, or a stronger safety net. They claim taxes are theft? Just who is stealing from whom here?Read more
Kansans Deserve Better
We never really thought it was over, did we? We suspected that the $500 million education finance legislation passed last weekend was not going to be enough to satisfy the Court. We expected we would have to revisit it. We just thought it would be in a Special Session this summer.
Now, word comes that the bill that was passed has an error in it. One that will short change many school districts to the tune of $80 million. Lawmakers will have to work extra fast to pass a correction when they return April 26, to meet the Court's deadline of April 30.Read more
Where are we now?
The Kansas Legislature has ended the the first, and most significant part of their session. They are scheduled to return on April 26, for the so called "veto session," although there are still several issues to resolve, among them a budget for next year, and the fate of the education amendment we talked about last week.Read more
Defending Education
Part of straddling the line between extremes is staying away from hyperbole. We work to encourage compromise and collaboration, and call out strident calls like "Taxes are theft!" for what they are. That said, we find we cannot emphasize this next point too much: education is under attack in Kansas, and it is time to stand up to defend it, lest we lose an entire generation of children.
This week in Topeka, hearings are underway on a proposed amendment to the Kansas Constitution to give control of education standards to the Legislature. If made law, this would irreparably hurt education in Kansas, and put one more nail in the coffin of the checks and balances that keep our government sound.
That sounds like a lot of hyperbole, right? Let us convince you.Read more
Arming Teachers is Ridiculous
In the wake of this weekend's worldwide outpouring of resistance to gun violence, and with a backdrop of little to no action on funding public schools, the Kansas Legislature is holding a hearing on a bill designed, not only to arm Kansas teachers, but to hold school districts liable should they not allow teachers to carry weapons and the unthinkable happens.
This is a terrible idea, and an incredibly tone-deaf move on the part of legislative leadership. We are stunned.
Nobody we have spoken to sincerely believes this bill (HB 2789) has the will behind it to pass, certainly not on the floor of either chamber if it makes it out of committee. So why is this bill being heard?Read more
ACTION ALERT - Oppose Anti-LGBT Adoptions Bills
The Kansas Legislature is hearing bills TOMORROW (Wed, 3/21/18) that would allow religious organizations to deny adoptions to LGBT families. In addition to the moral grounds (LGBT persons should not be discriminated against), and the data-driven grounds (LGBT families are at least as stable as heterosexual families), MainStream objects to this legislation because religious morality has no place in government.Read more
$2 billion
You may have heard that the Kansas Legislature on Friday received the report it paid for on the cost to adequately finance public schools in the state. You may have heard this because the numbers were eye-popping, between $400 million and $2.1 billion (with a B). It should be said that many expected the number delivered to be much lower. The report was commissioned by Republican leadership, already keen on spending less on public education. One of the people hired to deliver the report previously low-balled a similar report in Texas. The political rhetoric that surfaced this weekend has been pretty spectacular. And ridiculous.Read more
Education Movement Ahead?
Last week in Kansas politics had a few bright moments, but mostly it was still setup for the biggest issues still to come. While progress was made on stopping the Convention of the States, and Kris Kobach's defense of his citizenship requirement for voter registration faltered, we got muddled direction on gun safety from legislative leaders, and still no news on education finance, the deadline for which looms closer and closer.Read more


text:
Best Civil Contractors in Kolkata
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24 Civil Contractor in Kolkata
DZAYN STUDIOCircus Avenue, Lower Range, Ballygunge, Kolkata, West Bengal, IndiaHired 2 times on UrbanClap
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Creative GraphicKolkata4.2(6 reviews) Hired 3 times on UrbanClap
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- vverified5.0
Mr. Perfect !! He had great knowledge about work, he know how to make space in small place. Great design, great idea, and good person to hire... Tanks to urbanclap for keeping great professional.
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Excelent !! Modern moduler kitchen design.. I just love it...👍Read more reviews
Mandal Trading CompanyKolkata2.9(5 reviews) Hired 4 times on UrbanClap
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Projects and ServicesHowrah5.0(3 reviews)
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- aamiya5.0
We hired Chandranath and his firm for 5-6 projects of full office construction in different places like City Centre 1, Asansol, Sector 5 Kolkata etc. He quoted around 13Lakh for each project and the quality of work that he delivered was just amazing. He completes work right on time and is very good overall.
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We hired the firm for 2 projects one was for our home in Kastopur and other for our office in Krishnapalli. They did fantastic work on both the projects. They completed the work 1 day prior to finishing date and cost were 10k less than what they quoted.Read more reviews
DecormindAlipore, Kolkata, West Bengal, India2.2(3 reviews)
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- aananda4.5
My compliments to every person (especially the labourers) who carried out the renovations I was undergoing at home. Passionate, dedicated and worked above and beyond the call of duty - especially since I had a very tight deadline I was working towards. Were there a few errors? Yes. Did it go slightly over budget? Yes. But these are to be expected in any project, if they are not excessive. If I had to get further work done, I would definitely hire them again.
SP interiorGopalpur Gram, Kolkata, West Bengal, India4.5(1 review)
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- ddeblina4.5
Still now having a good experience. Will share the details after completion of work.
Amitava ChakrabortyBijoygarh, Kolkata, West Bengal, India1.0(1 review)
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Shree ConstructionRajpur Sonarpur, Kolkata, West Bengal, India5.0(1 review)
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Koushik MukherjeeAlaknanda Apartment, Jelepara, Surjaysen Nagar, Khardaha, Kolkata, West Bengal, India
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7CInteriorPanihati Municipality, Sukchar, Jaiprakash Nagar, Khardah, Kolkata, West Bengal, India
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The nitrate reductase of the hyperthermophilic archaeonPyrobaculum aerophilum was purified 137-fold from the cytoplasmic membrane. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, the enzyme complex consists of three subunits with apparent molecular weights of 130,000, 52,000, and 32,000. The enzyme contained molybdenum (0.8-mol/mol complex), iron (15.4-mol/mol complex) and cytochrome b (0.49-mol/mol complex) as cofactors. The P. aerophilum nitrate reductase distinguishes itself from nitrate reductases of mesophilic bacteria and archaea by its very high specific activity using reduced benzyl viologen as the electron donor (V max with nitrate, 1,162 s−1 (326 U/mg);V max with chlorate, 1,348 s−1 (378 U/mg) [assayed at 75°C]). The Km values for nitrate and chlorate were 58 and 140 μM, respectively. Azide was a competitive inhibitor and cyanide was a noncompetitive inhibitor of the nitrate reductase activity. The temperature optimum for activity was >95°C. When incubated at 100°C, the purified nitrate reductase had a half-life of 1.5 h. This study constitutes the first description of a nitrate reductase from a hyperthermophilic archaeon.
Nitrate serves as electron acceptor to many prokaryotic microbes that thrive under anaerobic conditions. Nitrate respiration occurs via two independent pathways, the denitrification pathway and the ammonification pathway (3). Nitrate is reduced sequentially to dinitrogen gas in the denitrification pathway, while ammonium is the product of the two-step ammonification pathway. The first reaction, in which nitrate is reduced to nitrite via the membrane-bound nitrate reductase, is identical in both pathways (3, 21). In general, the dissimilatory nitrate reductase is conserved among bacteria and archaea that have been investigated thus far. The enzyme has been extensively studied in mesophilic nitrate reducing bacteria such as the ammonifierEscherichia coli and the denitrifiers Paracoccus denitrificans, Pseudomonas stuzeri, Pseudomonas denitrificans, and others (5, 6, 11, 12, 16). The E. coli NarGHI enzyme is one of the best-characterized enzymes (3). The enzyme complex consists of three subunits (11, 16). The α subunit (NarG) has an M r of 145,000 and contains a molybdopterin cofactor at its active site, where nitrate is reduced to nitrite. The β subunit (NarH) has anM r of 58,000 and is the location of one [3Fe-4S] center and three [4Fe-4S] centers. Both the α and β subunits are attached to the cytoplasmic membrane by the 25,000-Da γ subunit (NarI). This polypeptide contains cytochrome b and functions to oxidize the menaquinol or ubiquinol of the quinone pool. Electrons are transferred from the quinol pool via the β subunit to the α subunit active site (3). Recently, nitrate reductases from several archaeal species have been described. While Haloferax volcanii contains a heterotrimeric enzyme complex similar to the bacterial dissimilatory nitrate reductases, the nitrate reductase from Haloferax denitrificans was purified as a heterodimeric enzyme possibly lacking the membrane anchor subunit (4, 9). In contrast, Haloarcula marismortui contains a homotetrameric nitrate reductase (20).
The archaeon Pyrobaculum aerophilum is the only hyperthermophilic denitrifier that has been characterized thus far (18). P. aerophilum was isolated from a hot spring in Italy and grows optimally at 100°C (18). Based on 16S rRNA gene sequence analysis, P. aerophilum was placed in the order Thermoproteales within theCrenarchaeota branch of the phylogenetic tree (18). In contrast to its strictly anaerobic, sulfur-respiring relatives, P. aerophilum can respire aerobically with 0.6 to 1% oxygen present in the gas phase. Alternatively, P. aerophilum can grow anaerobically by dissimilatory nitrate reduction, a catabolic feature not found among its close relatives (1, 18). Previous studies have established that growth of P. aerophilum is strictly dependent on the availability of tungstate in the culture medium (1). Tungstate, however, has been demonstrated to inactivate molybdoenzymes such as the nitrate reductase and formate dehydrogenase in mesophilic bacteria (8). In contrast toE. coli, where nitrate reductase enzyme function was abolished by the addition of tungstate to the medium, the enzyme was still active in P. aerophilum cultured with elevated tungstate concentrations (1). Thus, P. aerophilum appears to be adapted to an extremely thermophilic environment that typically contains elevated levels of tungstate (1). In this study we report the purification and characterization of the thermostable nitrate reductase from P. aerophilum.
Growth conditions. P. aerophilum was grown anaerobically at 95°C in a marine medium containing yeast extract (0.05%), peptone (0.05%), nitrate (10 mM), molybdate (0.5 μM), and tungstate (0.4 μM) as previously described (1). For routine cell transfers, P. aerophilum was cultured in 50 ml of medium in 125-ml anaerobic serum vials (1). For large-scale cultivation, P. aerophilum was grown in 70 liters of culture medium in a 100-liter glass-lined fermentor (Pfaudler). Cultures were harvested in late exponential phase by concentration with a hollow-fiber filter (A/G Technology) and subsequent centrifugation for 20 min at 16,000 × g . The pelleted cells were stored at −20°C. Typical cell yields were 35 g (wet weight) of cells per 70 liter of culture volume. Cells that were used for the preparation of the membrane fraction for absorption spectroscopy were grown in the absence of molybdenum and with 4.5 μM tungstate.
Purification of the nitrate reductase.Enzyme purification was performed under aerobic conditions and at room temperature unless otherwise indicated. Frozen cells were resuspended in 20 mM sodium phosphate buffer (pH 7.0) and broken by sonication (1). The membrane fraction was collected by ultracentrifugation at 100,000 × g for 60 min at 4°C. The membrane pellet was then resuspended in 20 mM sodium phosphate (pH 7.0). Membrane-bound proteins were solubilized by extracting the membrane fraction with 2% Triton X-100 in 20 mM sodium phosphate (pH 7.0) at a ratio of 6 mg of Triton X-100 per mg of protein. The mixture was stirred for 45 min and centrifuged at 100,000 × g for 60 min at 4°C to remove the insoluble fraction. The supernatant was applied to a 5-ml Hi-Trap Q Sepharose column (Pharmacia Biotech) equilibrated with 20 mM sodium phosphate (pH 7.0) containing 0.1% Triton X-100. After the column was washed with 15 volumes of the same buffer, nitrate reductase activity was detected in the flowthrough fractions. The flowthrough fractions were loaded onto a 5-ml hydroxyapatite column (Bio-Rad) equilibrated with 10 mM sodium phosphate buffer (pH 7.2) containing 0.1% Triton X-100. A 10 to 200 mM linear sodium phosphate gradient was applied, and nitrate reductase activity was recovered in fractions containing about 130 mM sodium phosphate. Fractions containing nitrate reductase activity were combined and concentrated with Centricon 100 concentrators (Amicon). The buffer of the concentrated sample was exchanged, using a PD-10 Sephadex G-25 M column (Pharmacia) for the buffer used for the subsequent chromatography column (i.e., 20 mM Tricine [pH 8.0] plus 0.1% Triton X-100). The final chromatography step was performed with a MonoQ HR5/5 column (Pharmacia), which was developed with a linear gradient of NaCl from 0 to 200 mM. Nitrate reductase eluted at about 100 mM NaCl. The purified enzyme was concentrated and stored at 4°C or, for long-term storage, at −70°C.
Enzyme assay.Nitrate reductase activity was determined with reduced benzyl viologen as the electron donor in an anaerobic cuvette assay as previously described (1). The assay was performed at 75°C unless otherwise indicated. One unit of enzyme activity is defined as 1 μmol of nitrate reduced per min.
Characterization of the pH and temperature optima.The pH and temperature optima for nitrate reductase activity were measured at the pH values and temperatures indicated in the figures.
Determination of temperature stability.The temperature stability of the P. aerophilum nitrate reductase was determined by incubating the enzyme (0.2 mg of protein/ml) in 10 mM Tricine (pH 8.0)–0.1. M NaCl–0.1% Triton X-100 at the indicated temperatures for more than 30 h. Throughout the experiment, samples were removed and assayed for nitrate reductase activity at 75°C.
Protein determination.The protein concentration was determined by the modified Lowry procedure for membrane-bound and detergent-containing protein samples (Sigma). Bovine serum albumin was used as the standard.
Absorption spectroscopy.The oxidized and dithionite-reduced absorption spectra of the purified enzyme were recorded on a Beckman DU 640 spectrophotometer as previously described (17). The amount of cytochrome b present in the enzyme preparation was based on an extinction coefficient for cytochrome b of 20 mM−1 cm−1 at 556 nm (13). The absorption spectrum of the P. aerophilum membrane fraction was recorded by an SLM Aminco DW 2000 spectrophotometer using cuvettes with a 1-mm path length. After the spectrum of the oxidized sample was recorded, anaerobic conditions were established by three cycles of evacuation and flushing with N2 gas. The sample was subsequently reduced with 1 mM sodium dithionite, and the spectrum was recorded after 3 and 6 min at room temperature to ensure complete reduction of the cytochrome-containing membrane proteins. The oxidation of the nitrate reductase cytochrome b was recorded immediately after addition of sodium chlorate at 3.3 mM (final concentration).
Molecular weight determination.The apparent molecular weight of the purified, denatured nitrate reductase was determined from the mobility of the protein in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (12.5% polyacrylamide). The molecular weight standards were bovine serum albumin (M, 66,000), chicken egg albumin (M, 45,000), glyceraldehyde-3-phosphate-dehydrogenase (M, 36,000), bovine carbonic anhydrase (M, 29,000), bovine pancreas trypsinogen (M, 24,000), soybean trypsin inhibitor (M, 20,000), and bovine milk α-lactalbumin (M, 14,200) (Sigma).
Metal analysis.The molybdenum, tungsten, and iron content was determined using inductively coupled plasma mass spectroscopy performed by the Soil and Plant Analysis Laboratory, University of Wisconsin—Madison/Extension, Madison, Wis.
Gel electrophoresis.Protein samples were diluted 1:2 with 10 mM Tris-HCl (pH 8.0)–5% SDS–10% β-mercaptoethanol–0.02% bromphenol blue and incubated for 30 min at 100°C. The protein samples (1 μg) were then separated on an SDS–12.5% polyacrylamide gel (Phastsystem; Pharmacia). Precast polyacrylamide gels and SDS buffer strips were purchased from Pharmacia.
Chemicals.Benzyl viologen, Lubrol, Thesit, and Brij 35 were obtained from Sigma Chemical Co. Triton X-100 was purchased from Boehringer. All other chemicals were of the highest purity commercially available.
Purification of the nitrate reductase.The nitrate reductase from P. aerophilum was purified from cells that had been grown anaerobically in a nitrate-containing medium supplemented with molybdate and a limiting amount of tungstate (i.e., 0.4 μM). The latter metal oxyanion concentration had been shown previously to promote high nitrate reductase levels (1). Since nitrate reductase activity had been previously localized to the membrane fraction, membranes were extracted using several different detergents. About 100% extraction of nitrate reductase activity from the cytoplasmic membrane was accomplished with 2% Triton X-100 at a ratio of 6 mg of Triton X-100 per mg of protein. A 5% solution of the nonionic detergent Thesit extracted 60% of the enzyme activity, whereas neither 5% Lubrol nor 5% Brij 35 extracted any nitrate reductase activity (data not shown). The detergents used did not aversely affect the nitrate reductase activity.
Nitrate reductase was purified from the Triton X-100 extract by Q-Sepharose, hydroxyapatite, and Mono Q ion-exchange chromatography (Table 1). The enzyme separated as a broad band on the last column. Only the peak fractions were combined to give the final preparation. This, however, resulted in a low yield (2%). After the last purification step, the nitrate reductase enzyme was enriched 137-fold with respect to the membranes.
The active nitrate reductase at the final purification step consisted of three polypeptides with apparent molecular weights of 130,000, 52,000, and 32,000 as determined by SDS-PAGE (Fig.1). The purity of the enzyme preparation was estimated to be about 80% as judged from SDS-PAGE. Based on the purity of the preparation and using the enrichment factor of 137, it was calculated that the nitrate reductase constitutes about 0.6% of the membrane-bound protein.
Cofactor composition.The purified nitrate reductase exhibited a visible absorbance spectrum characteristic of cytochrome b(Fig. 2, traces C and D), with absorbance maxima of the reduced enzyme at 420 and 555 nm for the γ and α band, respectively. Based on the calculated M, of 214,000 for the three-subunit complex (see below), the amount of cytochromeb was estimated to be 0.49 mol/mol of enzyme complex. Nitrate or the alternative substrate chlorate can reoxidize the reduced cytochrome b of nitrate reductases. This can be demonstrated with the nitrate reductase present in the membrane fraction of the denitrifying bacterium Paracoccus denitrificans by adding chlorate to dithionite reduced membranes (Fig. 2, trace A). Unlike nitrite, the product of nitrate reduction, chlorite, the product of chlorate reduction, cannot be further reduced by other cytochrome-containing enzymes of the denitrification pathway and will therefore not interfere with the difference spectrum. InParacoccus denitrificans, the absorbance maxima for the nitrate reductase cytochrome b in the reduced-minus-oxidized difference spectrum are identical to those for the purified enzyme (7). The same experiment performed with membranes ofP. aerophilum identifies a similar chlorate-oxidizable cytochrome b (Fig. 2, trace B). The spectrum of the chlorate-oxidizable cytochrome b in the membranes is very similar to that of the chlorate-oxidizable cytochrome b in the purified Nar (trace E). The spectrum is also similar to that of the spectrum shown in trace D, which was generated from the absorption spectra of purified, air-oxidized, and dithionite reduced nitrate reductase. This is conclusive for the existence of a chlorate-oxidizable cytochrome b in the P. aerophilum nitrate reductase.
To determine whether this nitrate reductase contains molybdenum or tungsten as a cofactor, the molybdenum content of the purified nitrate reductase was analyzed by inductively coupled plasma mass spectroscopy. The enzyme contained 3.6 nmol of molybdenum and 0.27 nmol of tungsten per mg of protein. Based on the M r of 214,000 for the nitrate reductase complex and not normalizing for the protein purity, 0.8 mol of molybdenum and 0.06 mol of tungsten per mol of complex can be calculated. Thus, the P. aerophilumnitrate reductase is a molybdenum-containing enzyme similar to the enzymes purified from mesophilic bacteria. Iron was present at 15.4 mol/mol of enzyme (72 nmol/mg of protein), consistent with the notion that the enzyme contains one [3Fe-4S] cluster and three [4Fe-4S] clusters, similar to mesophilic nitrate reductases (3,21).
Kinetic properties.Using reduced benzyl viologen as the electron donor and either nitrate or chlorate as the electron acceptor, the kinetic properties of the purified nitrate reductase were determined. The Km value for nitrate was 58 μM, with a V max of 1162 s−1 (326 U/mg). The affinity of nitrate reductase was weaker when chlorate was used as a substrate, i.e., a Km of 140 μM; theV max with chlorate was measured to be 1,348 s−1 (378 U/mg).
Both azide and cyanide, known inhibitors of dissimilatory nitrate reductases in mesophilic microbes, were tested for their effect onP. aerophilum nitrate reductase activity (21). Azide was determined to be a competitive inhibitor of the P. aerophilum nitrate reductase with a Ki value of 31 μM. Cyanide was a noncompetitive inhibitor, which reduced the V max of the enzyme by threefold (i.e., to 108 U/mg). The pH optimum of nitrate reductase was determined to be pH 6.5 (Fig. 3A).
Temperature optimum and stability.The P. aerophilum nitrate reductase exhibited its highest activity at or above 95°C (Fig. 3B), which is slightly below the optimal growth temperature of P. aerophilum (i.e., 100°C). Noteworthy, at 37 to 45°C, nitrate reductase still exhibited 10% of its optimal activity. The Arrhenius plot indicates an activation energy of 36.6 kJ/mol for nitrate reduction (Fig. 3B, inset).
After incubation of purified nitrate reductase at 100°C, its activity declined with a half-life of 1.5 h (Fig.4). In contrast, the half-life of nitrate reductase activity within the cell membranes was 6 h (data not shown), suggesting that the lipid environment stabilized the enzyme. At incubation temperatures of 85 and 65°C, the purified enzyme exhibited a t 1/2 of 16 and 32 h, respectively (Fig.4). The enzyme was stable at room temperature for many weeks, with negligible loss of activity.
Hyperthermophiles, such as the denitrifying P. aerophilum, are naturally exposed to high levels of tungsten, a heavy metal that is abundant in high-temperature environments (14). In E. coli, anaerobic growth with nitrate is abolished when tungstate is present in the medium. Growth inhibition is based on the inactivation of molybdenum-containing enzymes such as nitrate reductase by the incorporation of tungsten in place of molybdenum into molybdenum cofactor of the enzyme (8). Previous studies have shown that the hyperthermophile P. aerophilum is well adapted to a high-tungsten environment in that it has a strict requirement for this heavy metal for its anaerobic growth mode on nitrate (1). However, also in this organism, nitrate reductase activity is decreased when the tungstate concentration in the environment is increased, suggesting that this enzyme might contain molybdenum as a cofactor (1). In contrast to other mesophilic nitrate reducers, P. aerophilumgrowth with nitrate is not abolished at high tungstate concentrations.
The purpose of this study was to characterize the nitrate reductase from this organism and to determine the nature of its cofactors. This report is the first description of a thermostable nitrate reductase from a hyperthermophilic archaeon. The enzyme was purified from cells that had been grown anaerobically with nitrate and in the presence of low levels of tungstate and molybdenum. These growth conditions had been demonstrated previously to promote the highest nitrate reductase activity (1). We demonstrate here that the P. aerophilum nitrate reductase is also a molybdenum-containing enzyme (0.8 mol of molybdenum/mol of enzyme) with similar subunit composition to that found for mesophilic nitrate reducers (3,21). In addition, cytochrome b is an integral cofactor of the P. aerophilum nitrate reductase, which can be reoxidized by its substrate in a similar fashion to that shown for the mesophilic bacterium Paracoccus denitrificans (Fig. 2).
The P. aerophilum nitrate reductase distinguishes itself from mesophilic membrane-bound nitrate reductases by its unusually high specific activity of 326 U/mg as measured at 75°C with reduced benzyl viologen as the electron donor (Table 1). Considering that the activity at 75°C is 62% of that at 95°C, nitrate reductase activity under physiological conditions would be about 526 U/mg (turnover of 1,875 s−1). This is about 7 to 40 times higher than the activity of nitrate reductases from mesophilic bacteria and archaea measured at their respective optimal temperatures. Typically, enzymes isolated from hyperthermophiles, such as the NAD-dependent glutamate dehydrogenase from the related Pyrobaculum islandicum, have comparable specific activities to those of enzymes from their mesophilic counterparts (15). Like other mesophilic dissimilatory nitrate reductases, the P. aerophilum enzyme can also utilize chlorate as a substrate at a rate slightly higher than the nitrate reduction rate. Interestingly, the apparent affinities for both nitrate and chlorate are 4- to 60-fold higher than those reported for other dissimilatory nitrate reductases (e.g.,K m of 0.2 to 3 mM) (21). It is possible that this highly active nitrate reductase may be an adaptation to counteract inhibition by tungsten under physiological growth conditions: P. aerophilum needs to support growth by nitrate respiration even when the tungsten concentration in the environment is high.
As is typical for enzymes of hyperthermophiles, the P. aerophilum nitrate reductase is most active at or above 95°C. The enzyme appears to be stabilized by its membrane environment, since detergent extraction results in a fourfold loss of the thermostability of nitrate reductase activity.
Denitrification in the archaea is in general not well understood. Archaeal denitrifiers that have been isolated thus far include P. aerophilum, the hyperthermophilic Ferroglobus placidus, and the several Haloferax species (1, 2, 10, 18-20,21). Thus far, five nitrate reductases, including the enzyme described in this study and one copper-containing nitrite reductase, have been reported (4, 9, 20). In subunit composition, theP. aerophilum nitrate reductase appears to be most similar to the Haloferax volcanii enzyme, which also consists of three subunits with molecular masses of 100, 60, and 31 kDa (4). Like the P. aerophilum enzyme, theH. volcanii nitrate reductase is membrane bound. In contrast, soluble nitrate reductases were purified from Haloferax denitrificans, Haloferax mediterranei, and Haloarcula marismortui (2, 9, 20). It is possible that an additional protein exists that associates these soluble nitrate reductases with the cytoplasmic membrane. The Haloarcula marismortui enzyme appears to differ from all the other enzymes in that it comprises a homotetrameric complex composed of a 63-kDa polypeptide (20). All nitrate reductases from halophilic archaea except the enzyme from H. denitrificans require high salt concentrations for activity and structural stability (4). It is interesting that the presence of increasing salt concentrations shifts the temperature optimum for activity of theH. mediterranei enzyme to a higher value (2). Thus, temperature optima for nitrate reduction of 59 and 89°C were assayed in the presence of 0.6 and 3.2 M NaCl, respectively.
In conclusion, the nitrate reductase isolated from P. aerophilum is a molybdenum-containing enzyme with unusually high specific activity. From an evolutionary point of view, the enzyme fromP. aerophilum is the oldest nitrate reductase characterized thus far and argues for the presence of a heterotrimeric enzyme in the last common-ancestor group of microbes.
This work was supported by a grant from the National Science Foundation (MCB-9631006). Some of the recent work was also supported by grant MCB-0091351 from the National Science Foundation.
- Received 12 July 2000.
- Accepted 1 July 2001.
- Copyright © 2001 American Society for Microbiology


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Child Welfare
> CW204 Child Welfare II - Family Assessment and Family Centered Practice
CW204 Child Welfare II - Family Assessment and Family Centered Practice
This module focuses on assessments that form the foundation of effective practice with children and families. Family-centered assessment focuses on the whole family, values family participation and experience, and respects the family's culture and ethnicity. Family-centered assessment helps families identify their strengths, needs, and available resources to develop a service plan that assists them in achieving and maintaining safety, permanency, and well-being.
Course Objective
• Describe the purpose of family assessments.
• List the different types of assessments and the information gathered in each.
• Describe family-centered practice approaches and how they can be applied to the target population.
• Describe informal and formal assessment practices to discover the needs, desires, and interests of children and families.
• Identify strategies for integrating gathered assessment data into effective case plans with families.
Intended Audience
Direct support professionals, case managers, nursing assistants, health aides
Author Bio
The Massachusetts Council of Human Service Providers, Inc. also known as Providers' Council is a statewide association of health and human service agencies. Founded in 1975, the Providers' Council is the state's largest human service trade association and is widely recognized as the official voice of the private human service provider industry. Providers' Council founded and deployed the first online curriculum and credential programs for direct support professionals and continues to enhance this curriculum with current data and workforce practices.
1.0 Hour
1.00 CE Credit(s)
Other Courses You May Be Interested In
Child Welfare
CWS03 Child Welfare - Families of Murder
Families in which murder has occurred and how to treat.
Substance Use and Addictive Disorders
SU09 Substance Use and Addictive Disorders - Crisis Intervention and Emergency Services
The purpose of this module to introduce you to the skills and knowledge needed to ensure safety and well-being for the individuals you support, your colleagues and yourself. Potential crisis situations are identified. This training does not endorse any specific physical interventions.
Autism Spectrum Disorder
ASD09 Autism Spectrum Disorder - Crisis Intervention
This module is designed to help give you the knowledge and skills to ensure safety and well-being for the individuals with whom you work, your colleagues, and yourself. Crises occur in the work that we do. But you can learn to identify potential crisis situations and be prepared to help manage the situation.


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Old Car Drawing Easy Rusty Cartoon Cool
|Include In Article
Old Car Drawing
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|Wednesday , June 12th , 2019 - 00:14:33 AM
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Quote from Old Car Drawing :
Set the box plot grobs in the scatter plot. Proceed to the theatre to find a play, or read a different type of book to what you normally read. There are now effortless reference guides teaching you just how to resemble a pro, utilizing a marker. To create the ideal wheelchair, you don't have to be a specialist with perspective drawing. Without the appropriate internet advertising resources the site could never be known outside of some family members and friends, thus you might find that your web business struggles accordingly. For instance, imagine that you own a site that sells mediation supplies.
If this is the case, you're halfway there. Developing a comic can be both fun and rewarding, so let's look at the four steps you may take that will reveal to you the way to draw comics and make your very own comic strip. It is crucial to keep up a skilled and uncluttered appearance. You're able to have a look at all 3 tips utilizing the step-by-step video lesson below.
If you want more things to draw here's a list full of ideas. There are a lot of diverse kinds of cool cars you may pick. At last, you can finish the entire car so it is going to appear to be a true vehicle picture. Comparable to the preceding steps, you don't need to draw the actual look car at first.
Simple to draw and fun for everybody. All you have to do is have fun! With the right coloring, you are able to make your hand-drawn diamond really cool and enjoyable.
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The man who has helped veterans in Alachua County for more than two decades is retiring. The veteran community in Gainesville consists of 20,000 people. Jim Lynch has helped secure benefits for many of them. WUFT's Erica Cordell sat down with Lynch on the eve of his retirement to talk about the past twenty years on the job.[audio:http://www.wuft.org/media/audio/89LynchQA.mp3]
Home / Florida Good / Alachua County Veterans Service Director Jim Lynch Reflects On The Past Two Decades
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Diploma Dreams Become Reality For Inmates At Graduation
In 2016, the Florida Gateway College became part of the Second Chance Pell Pilot Program. Now, three years later, 47 inmates walked at the first college graduation inside a Florida prison.


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Postmodern Marketing
of work that involves continuous learning and endless innovation. A technologically innovative computer firm may be energized by the same Sage archetype motivations. Quite unconsciously, the management of these companies tends to be attracted to brand identities consistent with the archetypes that simultaneously are shaping their own behavior and the corporate culture. This is how some leading companies happen onto archetypal identities and manage to retain them over time—especially if they have leadership that trusts their own insides and their intuitive hunches. They like brand identities that are like them. However, if they don't—and if marketing firms convince them to follow every fad or public whim—they inevitably will drift from one identity to another, creating no clear lasting impression. Marketing and advertising firms also have their unconscious biases. Within the field, individuals and firms have quite different reasons for getting out of bed in the morning. We all know people in advertising who have a novel in the drawer or ideas for making videos or films. They are in advertising because the field offers a well-paid way to express their creativity and artistry. We also may know people who love the competitive aspects of marketing—and are propelled onward by their enjoyment of the contest. Still others like the mental stimulation of marketing strategy. In archetypal terms, these examples reflect the archetypes of the Creator, the Hero (as competitor), and the Sage. If people in the field are uneducated about archetypes, the approaches they try to sell may simply reflect their own unconscious predilections and not be optimal for the client. To determine the brand identity a company will like, it is best to find out who the firm thinks it is—in terms of the archetypes it is living out. The biographies of successful business leaders demonstrate that, generally, they are drawn to a field or a product for some reason. Even in a societal context, where money and success are primary, individuals' deeper values are reflected in the details of their ambitions—the dreams that propel them forward. These values and dreams create an identifiable organizational culture, which is then reflected in its brand identity (e.g., IBM vs. Apple). We can trace the values to the underlying archetypes (in this case, Ruler and Outlaw/ Rebel, respectively), making it possible to factor the archetype of the organization into discussions of brand identity.


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Kayak Noosa, Noosaville
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Trip Planner: Australia / Queensland / Sunshine Coast / Noosa / Noosaville / Kayak Noosa
The new team at Kayak Noosa's mission is to share the delights of paddling kayaks and stand up paddle boards on the beautiful Noosa River with people of all ages and abilities.Put Kayak Noosa at the forefront of your travel plans using our Noosa road trip planning app .
Specialising in guided kayak adventures, paddle equipment sales, freedom hire, coaching and group programs.
Our unique river beach location right beside the Boathouse restaurant is a great place to chill out and get into the paddling groove for fun and fitness.
Our award winning sustainable business practices highlight our commitment to the environment and the conservation of our precious marine ecosystems and wildlife.
Join one of our guided kayak adventures and experience the thrill of river and ocean paddling and kayak sailing in the amazingly beautiful Noosa region.
We provide all the gear, equipment and professional guides and instructors. All you need is your sprit of adventure!
Freedom Kayak Hire - Explore Noosa River in your own time
We make it easy for you to customise your own kayak adventure by making our fleet of kayaks available for hire. We call this service our Freedom Hire as it allows you the freedom to explore the local waterways at your own pace. Kayak Noosa offer the best range of sea kayaks, sit-ons and racing skis in fibreglass, carbon and polyethylene available in the Noosa region. Ask us about our extensive range of kayaks and stand-up paddle boards also available for hire.
Kayak Centre - Try Before You Buy
Our on-water shop is conveniently situated on the Noosa River at the Boathouse Marina at Noosaville. We stock the best sea kayaks, recreational sit-on kayaks and fitness skis, along with an extensive range of kayak and paddle accessories, that will have you on the water in comfort, style and safety.
We have always extended our Try Before You Buy policy and believe it's the only way to match you with the right, river or sea kayak and associated gear. We ensure all your requirements are met through our extensive industry knowledge, professional sales staff and quality service.
Last but not least, we are paddlers and coaches ourselves and can't wait to show you the delights of 'Paddling in Paradise'
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Kayak Noosa Reviews
Hidden stingrays, swims in crystal clear water, watching soldier crabs scurry along the beautiful white Noosa sand, paddling among the mangroves, reveling in the silence. This was such a wonderful thi... more »
We went kayaking with the girls from this business with a mixed group of young children, teenagers and our 70+ mother! It was the most fun the kids had on their summer holiday. The girls were so suppo... more »
Great experience. We left the busy Marina and kayaked the water paths. So peaceful.
What a great trip! Erica and Tracy are incredibly friendly and some of the most knowledgeable guides you could ask for. We saw wildlife, learned about the area and had a fantastic sunset all from some top of the line kayaking equipment. Highly recommended!
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7 days in Brisbane, Sunshine Coast & Gold Coast BY A USER FROM AUSTRALIA April, popular sights PREFERENCES: April ATTRACTION STYLE: Popular sights PACE: Medium 26 days in Australia BY A USER FROM CANADA March, outdoors, wildlife, hidden gems PREFERENCES: March, outdoors, wildlife ATTRACTION STYLE: Hidden gems PACE: Medium 3 days in Sunshine Coast BY A USER FROM AUSTRALIA February, slow & easy, hidden gems PREFERENCES: February ATTRACTION STYLE: Hidden gems PACE: Slow & easy 10 days in Noosa BY A USER FROM AUSTRALIA April, culture, outdoors, relaxing, romantic, shopping, wildlife, popular sights PREFERENCES: April, culture, outdoors, relaxing, romantic, shopping, wildlife ATTRACTION STYLE: Popular sights PACE: Medium 8 days in Sunshine Coast BY A USER FROM AUSTRALIA March, relaxing, beaches, historic sites, hidden gems PREFERENCES: March, relaxing, beaches, historic sites ATTRACTION STYLE: Hidden gems PACE: Medium
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Read online or Download Best Christmas Pageant Ever / Edition 1 (Full PDF ebook with essay, research paper)
by Barbara Robinson
The horrible Herdmans are the meanest kids around. They lie, steal, cuss, and smoke cigars -- even the girls. The last place anyone expected to see them is in church. So when the Herdmans storm Sunday school and take over the annual Christmas pageant, everyone braces for the worst. But no one is prepared for what really happens when the rottenest kids in the world take over the greatest story ever told. It's a pageant full of surprises for everyone -- including the Herdmans themselves.
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Move-in ready! This newly updated waterfront residence in Royal Palm Yacht & Country Club sits on 100' of water with a private dock, and direct Intracoastal and Ocean access. The home features 4 bedrooms, 4.5 baths and has been updated with new flooring, new kitchen and new baths. Open pool/patio area with great waterfront views and east of the bridges. Easy to show, and available to move into today!
Fireplace(s), Foyer, Split Bedroom, Walk-in Closet
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All listings featuring the BMLS logo are provided by Beaches MLS Inc. Copyright 2019 Beaches MLS. This information is not verified for authenticity or accuracy and is not guaranteed.
© 2019 Beaches Multiple Listing Service, Inc. All rights reserved.
Listing information last updated on June 24th, 2019 at 8:15am CDT.


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About this Event
All across the globe, there is increasing recognition of the transformative power of creativeplacemaking to revive the economic and cultural life of cities. Singapore is no exception. Since2008, the Singapore government has been engaged in a concerted effort to placemakeSingapore into a culturally-vibrant cityscape with "heart and soul". These efforts are part of anongoing whole-of-government strategy called "place management", which aims to enliven andtransform public spaces into people-friendly spaces that engender a sense of communityownership and connection. The arts have been identified as an expedient resource to enable thistransformation.
However, despite its increasingly global popularity as a policy buzzword, what constitutes successful creative placemaking and its associated outcomes remain vague and tenuous. Notably, scant critical attention has been paid to the spatial and social embedding of global urban cultural policy discourses in Singapore, particularly its impact on the localised dynamics of arts spaces and practices.
Using Singapore as a case study for provocation, this session aims to stimulate critical discussionon the emergence of creative placemaking as an urban cultural policy and the ensuingimplications for local arts practice. Through a granular and situated analysis of local cultural policy,ethnographic and visual material from Singapore, this session will consider both the culturalcontradictions and redistributive potentials of a state-driven approach to creativeplacemaking/management. Ultimately, this session questions how we can better understand andaccount for the impact and interplay of global policy transfer and mobility, particularly in relationto site-specific conditions of artistic practice and cultural production.
Speaker: Dr. Su Fern Hoe (Singapore Management University)
Respondent: Dr. Tarek Virani (Queen Mary University of London)
Chair: Dr. Jenny Mbaye (CCCI, City, University of London)
The Centre for Culture and the Creative Industries (CCCI) at City, University of London presents a critical conversation about art spaces and creative clusters in Singapore.
This event is part of the CCCI event series spreading throughout May and early June to celebrate the UN World Day of Cultural Diversity for Dialogue and Development (May 21st).
Find more about the series here: https://www.city.ac.uk/news/2019/may/centre-for-culture-and-the-creative-industries-to-curate-event-series
About Dr Su Fern Hoe:
HOE Su Fern is an arts researcher, educator and coordinator who traverses artistic disciplines and mediums. She is currently Assistant Professor and Programme Leader of Arts and Cultural Management at the Singapore Management University. She holds a PhD in Culture and Communication from The University of Melbourne. Her research areas include arts and cultural policy studies, urban cultural economies, arts spaces and placemaking. Her research is informed by her pursuit for practice-oriented and engaged arts research and her interest in enhancing research impact beyond academia, particularly through the power of the arts to catalyse dialogue and bridge differences.
About Dr Tarek Virani:
Tarek Virani's research spans a number of areas within the creative and cultural economy including: the role of knowledge in the creative and cultural economy, cultural policy, artistic knowledge within locally bounded artistic communities, new work spaces in the creative and cultural economy, the role of microcommunity engagement in culture-led regeneration, the role of the creative and cultural economy intermediary, the link between the creative industries and local development, creative and cultural hubs, and the internationalisation of creative work. Tarek has done work for a number of local, national and international organisations including research institutions and local authorities.
He is Lecturer of Creative Industries at the School of Business and Management (SBM) at Queen Mary University of London where he teaches research methodologies and the creative industries at both undergraduate and postgraduate levels. Finally, Tarek is co-convener of the new MA in Creative Industries and Arts Organisation at SBM.
About CCCI event series 2019:
From Senegal to Singapore: Creative Spaces in a Globalised World
The series of events gathers a range of international experts from across the fields of culture and creative industries to showcase and share their experiences of developing spaces for arts and creativity in their countries.
Bringing together experts and the public, these events will be reflective and interactive spaces to explore and exchange ideas on areas such as culture, policy and management.
If you have an interest in how creative spaces are being carved out in a globalised world then these events offer a great chance to hear stories from all around the world, as well as network and gather with peers.
This event is open to the public and free to attend however, booking is essential.
For more information, please contact: firstname.lastname@example.com
Dr. Su Fern Hoe (Singapore Management University)
Dr. Tarek Virani (Queen Mary University of London)