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TORONTO, ONTARIO–(Marketwired – June 7, 2016) – Rio Novo Gold Inc. (TSX:RN)(TSX:RN.WT) ("Rio Novo" or the "Company") announces that it has engaged Runge Pincock Minarco ("RPM") (formerly Pincock Allen & Holt) to update the results of the definitive feasibility study (the "2013 FS") for Rio Novo's 100%-owned Almas gold project ("Almas"), located in Tocantins State, Brazil, which was published in 2013.
In order to update the 2013 FS, Rio Novo senior management has engaged RPM (which also authored the 2013 FS) to examine how changes in the price of gold, as well as changes in the Brazilian mining industry and foreign exchange markets may have affected construction and operating costs. Results of the updated study are expected in the 3rd quarter.
RPM is Rio Novo's independent engineering consultant, and the RPM personnel who will author/supervise the updated study will be "qualified persons" under National Instrument 43-101 Standards of Disclosure for Mineral Projects.
About Rio Novo
Rio Novo is focused on the acquisition, exploration and development of gold properties in Brazil and Colombia. The Company's short-term goal is to become a producer of gold by bringing Almas, located in the State of Tocantins, into production. Almas enjoys both established infrastructure and main grid hydropower in a proven and mining friendly jurisdiction and is permitted for construction.
This press release contains forward-looking statements. All statements, other than of historical fact, that address activities, events or developments that the Company believes, expects or anticipates will or may occur in the future (including, without limitation, statements regarding the estimation of mineral resources, exploration results, potential mineralization, potential mineral resources and mineral reserves) are forward-looking statements. Forward-looking statements are often identifiable by the use of words such as "anticipate", "believe", "plan", may", "could", "would", "might" or "will", "estimates", "expect", "intend", "budget", "scheduled", "forecasts" and similar expressions or variations (including negative variations) of such words and phrases. Forward-looking statements are subject to a number of risks and uncertainties, many of which differ materially from those discussed in the forward-looking statements. Factors that could cause actual results or events to differ materially from current expectations include, among other things, without limitation, failure to establish estimated mineral resources, the possibility that future exploration results will not be consistent with the Company's expectations, the price of gold and other risks identified in the Company's most recent annual information form filed with the Canadian securities regulatory authorities on SEDAR.com. Any forward-looking statement speaks only as of the date on which it is made and, except as may be required by applicable securities laws, the Company disclaims any intent or obligation to update any forward-looking statements.

subset: commoncrawl


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We have just come back from Texas School 2014. We have another week of education under our belts. Please call or come by to schedule your next portrait session.
The above photo is some of the friends at Texas School 2014, some old, some new but all of us share a love of photography.
The above image was taken during the Thursday night Shootout during Texas School ! She is a model doing a steampunk style of clothing.


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2018 NBA All-Stars - Having two All-Stars shows how far the Minnesota Timberwolves have come
Until this year, the Minnesota Timberwolves had only ever had five All-Star players in the history of their franchise, and only three that were not named Kevin. Messrs Garnett and Love account for 13 of the team's 16 all-time All-Star appearances, the other three being one-off appearances for each of Sam Cassell (2004), Wally Szcerbiak (2002) and Tom Gugliotta (1996).
Only one Wolves player has been an All-Star since 2007, that being Love, who made it for three out of the four seasons between 2010/11 and 2013/14. At the time of writing, the franchise has not made the playoffs since that season, nor even a couple before it, last making the postseason via a Western Conference Finals run back in 2003/04. That season was also the most successful one in franchise history; it, plus the seven consecutive first-round exits in the seven seasons immediately prior, and the short list of All-Stars above, is all the success the franchise has ever enjoyed. And 'success' here is a very relative term.
That, however, is about to change. The Timberwolves are headed back to the playoffs this season, and are doing so behind the fearsome duo of Jimmy Butler and Karl-Anthony Towns.
The years of treading water after the departure of Kevin Garnett saw the Timberwolves slowly accumulate a slew of young players and tradeable assets. The quality of these assets and players, however, never usually quite caught up to the quantity.
With some big misses along the way - drafting Jonny Flynn ahead of Stephen Curry in 2009; trading the pick that could have been Ricky Rubio (or Curry again) for Mike Miller that same year; drafting Wesley Johnson fourth in 2010 ahead of DeMarcus Cousins; trading the pick that could have been Eric Bledsoe for Martell Webster; picking Derrick Williams second in an admittedly dry draft year in 2011; taking Shabazz Muhammad and Gorgui Dieng ahead of Giannis Antetokounmpo and Rudy Gobert in 2013 - the Wolves never got the elite quality that would break the cycle of mediocrity. Love was somewhat of the exception, but, as above, not even he could drag the team to the playoffs.
The cycle was finally broken, however, with the drafting of Towns.
The Wolves were perversely lucky to be so bad in the 2014/15 season (16-66) so as to be able to draft first overall. With that, they were in a position to draft Towns, who became the second-best player in franchise history pretty much from day one.
Offensively, Towns has it all. With touch, feel and an increasing amount of shooting range (via a flat-footed jump shot that takes a couple of weeks to get away), Towns is an elite big man scorer, averaging 20.1 points per game on a 64.7% true shooting percentage thus far this season. He runs, rolls, posts and spots up, floats, hits cutters, and has improved his scoring efficiency every season concurrent with lowering his turnover rate.
Towns's shot chart is a thing of beauty. He shoots 71.8% from the rim, 46.6% from three to ten feet, 46.5% from ten to sixteen feet, 50% from sixteen feet to the three-point line, and 42.9% from three. Those are above-average marks in every area. He arrived in the league as already a pretty complete offensive player, and yet he has only gotten better. On one end of the court, he is already about as good as it gets.
The defensive end is where Towns must now make the biggest strides. Towns frequently misses rotations, both on the perimeter and when he is supposed to be helping from the weakside on the interior, and whenever he misses these rotations, he does not put forth enough energy to try and recover to get there. When Towns is in the right position defensively, be it by chance or by a good rotation, he can impact the play. His long arms combine with genuinely good shotblocking timing see him generally make good decisions, save for going after some blocked shots that he shouldn't. But if he is not in the right position to begin with, he won't get there.
This one area is where Towns shows his youth and relative experience. Conversely, however, Jimmy Butler already does it on both ends.
Entering the league as an athlete, rebounder and somewhat disruptive defender, Jimmy Butler's upside was thought to be somewhere between Jarvis Hayes and Bonzi Wells. To get to any levels beyond that would have taken a development in his skills and poise on the ball that is almost never seen in one already with four years of college basketball already under his belt. Yet that is exactly what has happened.
At this point in his career, Butler is as confident on the ball as any wing player. He drives, pulls up or just raises up to shoot with a confidence and a swagger that rivals anyone, and with a nous and willingness to draw contact that is greater than most. Butler averages 22.5 points on only 15.8 shots per game, a remarkable 59.7% true shooting percentage from a star wing player born out of his 48% overall shooting from the field, 37.9% from three-point range and 86.2% free throw shooting on a striking 7.1 attempts per game. All the while, he remains the team's primary option for defending opposing stars, be they guards or big forwards.
For comparison's sake, his fellow starter on the wing Andrew Wiggins is taking the exact same number of shots per game, but scoring only 17.9 points per contest, and not being as entrusted, effective or versatile defensively as Butler. Butler is playing at an MVP calibre level, just like he was last year. This year, however, he is on a team good enough for people to notice.
The quantity of young assets that had given the Timberwolves an often anticlimactic hope for so long were finally consolidated in the trade for Butler over the summer. Zach LaVine, Kris Dunn and the 2017 #7 pick that was eventually used on Lauri Markkanen were moved for Butler and the #16 pick (used on Justin Patton). It was a statement of intent, and, rather than trading a trade for an aging Mike Miller or Marko Jaric or any other Wolvesian move of years past, a strong push forward.
And it is working.
Between Towns and Butler, the Timberwolves are headed back to the playoffs. They sport a 35-24 record, good for the fourth seed in the Western Conference, coming from a 113.8 offensive rating. That number is third in the league, and while head coach Tom Thibodeau has yet to impart his pioneering ICE-defensive schemes into his team (the 110.5 defensive rating is sixth worst in the league), the Timberwolves are firmly in the Western Conference's second tier.
The team blends youth and experience, with quality at every position. Lack of depth and concerns about Thibodeau's excessive minutes distribution notwithstanding, the Timberwolves are much improved and still improving.
And it all stems from having two 2018 NBA All-Stars in Jimmy Butler and Karl-Anthony Towns.News Now - Sport News


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Congratulations! Your promo code PROMOCODE will be automatically applied when you check out – enjoy.
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Music for Seasons
Music is the string that binds us to moments in time. Where in time am I now?
Music is the string that binds us to moments in time. Where in time am I now?
It's that time of year again, when musicians and music fans in Madison (and beyond!) votes for our favorite local acts. Voting is now closed, winners will be announced at the awards ceremony on June 9th! Thanks for supporting SNR and the MAMAs! Here are the 2019 nominees from the Sunday Night Records crew: Electronic … + More
You heard me. Look here: SNR #winning Cellist of the year: Allison Lenz Clarinetist of the year: Hannah Edlén Accordion of the year: Nate Meng and The Stolen Sea (amazing) Unique Performer award: Nate Meng and The Stolen Sea
Instructions Go to http://www.broadjam.com/mama/ If you have already registred, login and follow the instructions to vote. If not, click "fan membership." Fill out the form fields and check boxes. On this page you will be asked to make a $5 donation to the MAMAs. Money will fund school music programs in Madison and Dane County! Once registered, … + More
Andrew Christensen has been living and working in Bangor, Maine the last few months. He is returning January 5th for the release of his debut album, "Tomorrow I'm Gone", at the North Street Cabaret in Madison.. Below is a letter SNR's quarterback Ryan Lansing wrote to Andrew this summer, while he was finishing up the … + More
Y'all. Frandú. I first met Frandú at the Tip Top open mic about a year and a half ago while visiting Madison. He's a little ball of bright-hot energy bouncing around between the sky and the earth at the speed of light, a powerful presence in a small package that one can't quite seem to … + More
2017 is at last upon us, and SNR is already in full swing. Our 2017 releases are slated to begin in February, and include the considerable talents of Andrew C. , Chloe Louise, Frandu Smith, The Werewolverine, Nate Meng & The Stolen Sea, and All Other Names. We're still getting all our legal ducks in a … + More
Whether tears of holiday joy are freezing to your cheeks up in Madison, or you're having a Wet Christmas like I am out here in lala, we wish you all the love and warmth in the world. May your days be merry and musical, and may 2017 not be as much of a hot mess … + More
Welcome to the new SNR website. Sunday Night Records is a label out of Madison, WI. We're currently a three-man team consisting of Nate Meng (Artist Development: Madison); Ryan Lansing (Recording Engineer: Madison); and Jess Estes (Admin & Design: Los Angeles). Our collective credits include multiple records and tours. We are currently assembling our Listening Advisory … + More


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Research Repository
Overtraining During Preseason: Stress and Negative Affective States Among Professional Rugby Union Players
Nicholls, Adam, McKenna, Jim, Polman, Remco and Backhouse, Suzanne (2011) Overtraining During Preseason: Stress and Negative Affective States Among Professional Rugby Union Players. Journal of Clinical Sport Psychology, 5 (3). pp. 211-222. ISSN 1932-9261 (print) 1932-927X (online)
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The aim of this study was to explore the perceived factors that contribute to stress and negative affective states during preseason among a sample of professional rugby union players. The participants were 12 male professional rugby union players between 18 and 21 years of age (M age = 19 years, SD = 0.85). Data were collected via semistructured interviews and analyzed using an inductive content analysis procedure. Players identified training (structure and volume), the number of matches played and the recovery period, diet, sleep, and travel as factors that they believed contributed to their experience of stress and negative affective states. The present findings suggest that players may require more time to recover between matches, alongside interventions to help players manage the symptoms of stress and negative affect during times in which players are overtraining.
|Item Type:
|Uncontrolled Keywords:
|ResPubID23821, rugby union, training, performance, qualitative, interviews, affect
|Historical > Faculty/School/Research Centre/Department > Institute of Sport, Exercise and Active Living (ISEAL)
Current > FOR Classification > 1701 Psychology
Historical > SEO Classification > 970117 Expanding Knowledge in Psychology and Cognitive Sciences
|Depositing User:
|Date Deposited:
|24 Aug 2012 02:30
|Last Modified:
|21 May 2020 07:27
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OTTAWA, Canada -- Pleora Technologies, a globally trusted development partner for standards-based camera interfaces, is pleased to announce that its iPORT embedded camera interfaces have been chosen by PHOTONIS to extend its NOCTURN family of digital low-light CMOS cameras to provide GigE Vision and USB3 Vision connectivity.
"Our development efforts are a direct result of immediate demand from our customers", says Gregg Bell, President and CEO, PHOTONIS USA. "Pleora has a strong reputation as an experienced and high-value development partner. By using their USB3 Vision and GigE Vision embedded camera interfaces we can rapidly achieve compliance with those standards".
Pleora is now providing iPORT NTx-U3 embedded camera interfaces to manufacturers of industrial cameras and other imaging devices, allowing these companies to rapidly create products which are compliant with the newly-released USB3 Vision standard. End users of these imaging devices will benefit from rapid software application development, as the Pleora eBUS SDK version 4.0 has been designed to incorporate USB3 Vision and GigE Vision functionality into a single API.
"In developing the USB3 Vision compliant iPORT NTx-U3 embedded camera interface we took extraordinary efforts to ensure seamless mechanical, electrical and software compatibility between it and our latest generation of GigE Vision embedded camera interfaces", says George Chamberlain, President of Pleora Technologies. "This compatibility will help PHOTONIS to efficiently manufacture, maintain, and support the new designs, while giving their customers the ability to support both GigE Vision and USB3 Vision models using a single software library."
Exhibition at SPIE DS+S
Both PHOTONIS and Pleora Technologies will be exhibiting at the upcoming SPIE Defense, Security, and Sensing 2013 trade show beginning April 30 at the Baltimore Convention Center, Baltimore, Maryland. Visit PHOTONIS at booth #1941 and Pleora Technologies at booth #1242.
About Pleora
Pleora Technologies Inc. pioneered the use of Gigabit Ethernet for real-time camera connectivity and revolutionized industrial imaging. With this spirit of innovation, Pleora continues to engineer standards-based camera interface solutions for mission-critical imaging applications. Pleora's platform-based design approach offers a wide range of off-the-shelf and customized hardware and software components that provide maximum design flexibility while simplifying integration, maintenance and interoperability.
About PHOTONIS
PHOTONIS is a multinational high-technology group, with more than 40 years experience in manufacture, sales and innovation, specializing in photo sensor technology. The Group operates internationally in the night vision, surveillance, industrial, scientific, nuclear and medical imaging markets and celebrated its 75th anniversary in 2012. PHOTONIS USA holds several patents for the design and manufacture of a wide range of products including cameras, power tubes and photon detectors as well as standard and custom assemblies for mass spectroscopy and other medical, bio-medical and scientific instruments.


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The Liberal Party of New York is a unique type of political organization. Nothing like it exists anywhere else in the United States.
Despite this uniqueness, an understanding of the Liberal Party can be very helpful to anyone who wants to understand American labor politics at mid-century, precisely because this party exhibits in a striking and harshly developed form many of the characteristics and trends which exist in the rest of the field of labor politics in a less clear-cut way.
The Liberal Party is and has been since its inception a bundle of paradoxes and contradictions. It is a party based on labor which hotly denies that it is a labor party.
It is a party, furthermore, which represents only one section of the New York labor movement and is treated at best with coolness and usually with hostility by the rest of the labor movement.
It is an organizationally independent patty which seeks at most elections to transform Itself into an adjunct of the Democratic Party, and even when it is rebuffed and forced to run its own candidates, tries to disguise itself and its role as much as possible from the public.
It is a party which seeks to recruit members and activists on the basis of claims to its independence, incorruptibility and forward-looking, principled program; but when election-time rolls around, seeks to ram down the throats of these same members and activists one political deal with the Democrats after another, winks at the corruption of its ally, and tends to forget its program until after the election is safely past.
How has the Liberal Party managed to survive, much less to function all these years, in view of the fact that it is a sort of half-labor half-party? The answer is that it could only have survived this long because it has expressed in its own way the undecided, tension-ridden political position of American labor as a whole, and beyond that because it has expressed it in the peculiar political surroundings of New York.
The Liberal Party was originally created for the purpose of corralling the large radical and independent vote in New York for New Deal Democratic candidates. It was created by a section of the labor movement (the leaders of the International Ladies Garment Workers Union and the United Hat, Cap and Millinery Workers) with an old socialist tradition which had adapted itself in the main to the political philosophy of the American labor movement, but which sought an instrument through which to express and at the same time control the radical and independent sentiments and aspirations of the most advanced political-public in the country.
It is, on the one hand, a vestige of the socialist training and tradition of the older stratum of the radical wing of New York labor, and on the other an attempt to find a road to the independent political influence, action and power of the American labor movement of tomorrow.
Some examples of the twists and turns of Liberal Party politics during the past seven-years may be the-best way of illustrating haw the Liberal Party leadership has attempted to meet the dual pressures upon it: the -demand of Its active membership for more independence, and the acceptance by most of the membership and certainly by the leadership of its role as an adjunct of the Democratic Party.
In 1949 there was a New York City mayoralty campaign. The Democrats (backed by the bulk of the AFL and CIO) ran the incumbent Mayor O'Dwyer for re-election. The Liberal Party supported Newbold Morris in a coalition with the Republicans on a "clean government" basis.
Within one short year, the picture had changed completely. O'Dwyer resigned amid a big political stink. A Democratic hack, Impellitteri, ran as an Independent against the machine. The machine, in an effort to recover from the O'Dwyer scandal, nominated Judge Ferdinand Pecora for mayor, and the Liberals supported him.
Why? Because there was a gubernatorial campaign that year, and a senatorial one to boot. In their desire to re- elect Senator Lehman and to defeat the Dewey machine on a state level, the LP leadership decided to back the Democratic slate up and down the line.
The result: Impellitteri won a smashing victory against the New York Democratic machine, Lehman won handily, and for months the leadership had to explain to the membership why it had been necessary for them to exert themselves on behalf of a couple of party hacks like Pecora and Walter Lynch, the Democratic candidate for governor.
In 1951 the Liberal Party reached the peak of its prestige. Since there was no national or state campaign that year, the pressure of the ranks for an independent candidacy in the election for president of the City Council became overwhelming. The LP ran Rudolph Halley against candidates of both the Democrats and Republicans, and elected him in a smashing victory.
But in this campaign, as in all others, the party exhibited a marked reticence about appearing before the voters under its own name. It sought to set up an additional party in the hope of winning independent Republican votes, and won almost none. It permitted Halley to run his campaign on a straight anti-crime, anti-corruption basis, and put its city program in mothballs for the duration.
Nevertheless, the ranks and the leadership felt enormously encouraged by their success. It had been demonstrated that in certain circumstances the Liberal Party could be the strongest party in the city.
In 1952 the party leadership made a frantic attempt to use their new-found prestige to induce the Democrats to run acceptable candidates for major posts. Needless to say, on a national level they supported the Stevenson-Sparkman ticket. But the Democrats refused to nominate a liberal Democrat (like Harriman) for the Senate, and with obvious reluctance the LP leadership nominated their own candidate George S. Counts for the post.
He ran against liberal-Republican Ives and Democratic hack Cashmore and gained a whopping 485,000 votes on the Liberal Party line in the midst of the Eisenhower landslide. At the same time Stevenson got 410,000 votes on the Liberal line as compared to 220,000 for Truman in 1948.
Here again, a measure of independence, forced on a reluctant LP leadership, had paid off. In this campaign, again, the leadership had gone all-out for Stevenson, and had done very little to promote the Counts campaign. It was clear that a large number of voters want an independent party, and will show it whenever they are given half a chance.
In 1953 the New York City mayoralty campaign was again divorced from state and national campaigns. Early in the year, both Democratic and Republican Parties showed great division and uncertainty with regard to their candidates. After a long series of negotiations with both, the LP leadership found itself unable to make a satisfactory deal, decided to yield to the practically irresistible pressure from the ranks, and ran Halley again. The Democratic machine was thrown into panic by the prospect of losing City Hall outright, and got behind a reliable organization liberal, Robert Wagner Jr.
The campaign again showed the basic strength of the sentiment for independent politics in New York City. The LP got 468,000 votes while the Republicans, flushed with national victory, got 661,000, and Wagner was swept into office with over a million votes.
No campaign showed more clearly the distance between the LP and the rest of the labor-liberal movement in America than the campaign of 1963. Wagner was backed not only by both the AFL and CIO councils in the city, but by a whole galaxy of top national liberals from Mrs. Roosevelt up and down the line. Their cry was for party regularity, for sup- port to the Democratic Party as such.
Instead of reacting forthrightly and militarily to this challenge, the LP leadership sought to win by clever maneuver. Once again the campaign concentrated on "clean government" issues and virtually ignored the big economic and social problems of the city. Once again the LP sought to evade the challenge that it was ran by "labor bosses" by effacing itself during the campaign and disappearing into a series of "independents for Halley" organisations, with the idea that this was the way to lure independent votes into the fold.
The defeat of Halley for mayor seemed to take the starch out of the Liberal Party leadership. In 1964 they endorsed the whole Democratic slate in advance in exchange for the measly plum of Democratic endorsement for one Liberal for municipal judge.
Rumors flew thick and fast to the effect that the party was going to be dissolved. David Dubinsky, head of the ILGWU, openly speculated on whether the party should continue to exist, and informed it that it would have to count less heavily on the ILGWU for financial support than in the past.
The result was that Harriman, though elected governor, polled only 264,000 votes on the Liberal line . . . less than the unknown and defeated Lynch four years earlier.
In 1955 the LP annual dinner was dominated by Democratic dignitaries. Harriman and Wagner sat on the dais, along with De Sapio, the leader of Tammany Hall! Adolph Berle, state chairman of the party during the preceding years, resigned without bothering to wait till a successor had been found.
And it appears, at the present writing, that in 1956 the LP will just go through the motions while supporting Democrats at every level.
Flushed with the Halley victory in 1951, Alex Rose, a Liberal Party leader and president of the hat workers union, had written in his union's paper:
"The heads of central labor bodies across the land still play the game the old way, by way of promises of 'favors' from local bosses, by personal maneuvering with old-line politicians. They show no confidence in their own latent strength. They are either too lazy or too uncomprehending to assert the political power of their membership. With tragic consequences to the best interests of labor . . . they fail to mobilize all of labor's political potentialities."
The sad fact is that Rose and his colleagues in the leadership of the LP also "play the game the old way," though on a higher level than the men he condemned in the above paragraph.
They are not interested in the "personal" deals and maneuvers with old-line politicians in the narrow sense of the word "personal." But their whole method of politics is to seek to organize a mass base for the purpose of being able to exert pressure on the "old-line politicians," not for the purpose of winning elections and taking over the city and state governments.
Though they bemoan their isolation in New York, and tend to blame some of worst deals they have been forced to make on the fact that the rest of the American labor movement has failed to follow their example, they have not raised their voices in the national councils of labor to demand that the Liberal Party be extended nationally, or that a new party be formed into which it would be incorporated.
As a matter of fact, it is clear that the particular brand of politics of the Liberal Party can only continue to exist as long there is a real political hanger for an independent, honest party of the common people which is never satisfied. But while it feeds on this hunger, the LP leaders' brand of politics often tends to demoralise it rather than to satisfy it.
As year after year the ranks have to accept the leaders' stories of "great victories" won behind closed doors; as year after year they are compelled to support candidates whom they had previously opposed, or vice versa, they either turn into political cynics or leave politics altogether. There is a large turnover in the active membership, and a tendency for the old cadres to become bored, indifferent and inactive.
In addition, the LP leadership is now confronted with the fact of a united movement. That unity is bound to have consequences in the political functioning of the labor movement in America goes without saying.
To the extent that the LP is a unique maverick political animal, there will be pressure to kill it off and coordinate the politics of the unions which have been the backbone of the LP with those of the rest of the movement. Thus we may find that all the paradoxes and contradictions which have wracked the LP from its inception may be capped by the biggest and most ironical paradox of all: that the party fades just on the eve of the political realignment in America which it had so long awaited.
Fortunately, the LP is not foredoomed by any iron law of history to continue to waste away on the diet of crumbs from other parties' tables to which its leadership has confined it for so long. For in addition to this leadership, which is pretty well frozen in its hard-bitten, dogmatic theory of maneuverism, there has always been a sizable and healthy section of the rank and file which has pushed for a truly independent role for the Liberal Party."
They are convinced that the LP can actually win elections if it goes to the voters in its own name, with its own candidates, and advocates its own program during elections as well as in its educational literature.
And they understand that even if the party should fail at first to win as a really independent political movement, the tactic of running its own slate would put far more pressure on the other two parties to match it with good candidates of their own than has been the case when the LP has maneuvered for an acceptable candidate at the top of the ticket, and in return for this great boon from the Democrats, has agreed to support almost any machine hack they might run for all the offices lower down.
The pressures in American society which have brought about the unity of the labor movement are working inexorably to bring about a general political realignment also. By the very fact of its existence as a separate political party, the LP has been able to exercise more pressure on the Democrats in New York than has the labor movement in any other part of the country.
Though its successes have been minimized by the timid tactics of its leadership, once new political winds begin to blow the example of the Liberal Party will not go unnoticed by the militants in the rest of the labor movement throughout the country.
There is no denying the fact that the LP militants who are for an independent policy for the party have been wearied and in part demoralized by the tactics of their leaders. But the worst thing they could do now would be to give up the good fight.
Even though they have not yet succeeded in getting their party in any consistent way to play the inspiring role of which it is capable, their efforts have not been in vain. Compared to the political role of the bulk of the labor movement and the liberals in the country, theirs has been a noble one. And it may well be that the day is not far off when they will be able to set a real example, to spread the idea of truly independent labor politics far beyond the borders of New York.


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This is part 2 of a series that began with Functional Web Programming, Part 0 — Introduction.
If you haven't already, read Functional Web Programming, Part 1 — First-Class Functions.
What are pure functions?
In functional programming languages, (most) functions are pure. Pure functions have two really important traits:
- No side effects. Pure functions don't affect any part of your program outside of themselves — they can only work with data you pass to the function. They can't change that data, but can and usually do spit out remixed copies. Pure functions can't change program state; in fact, functional programs ideally are stateless (more on this in later posts).
- Always return the same value given the same input. Like the mathematical functions that are functional programming's namesake, pure functions always return the same result when fed the same data. If I have a function that returns 10 when I pass it 2, it will always return 10 when passed 2, no matter what.
Pure functions are referentially transparent, meaning that you can replace a call to a function with its output and not screw up the behavior of your app. This gives us some slick benefits, which I'll talk about in a minute.
First, though, let's peek at some examples of pure (and impure) functions in CoffeeScript and PHP. After that, I'll explain why you should try to write pure functions instead of impure functions in your program whenever possible.
Examples of Pure Functions
Here's an example of a really simple pure function, which returns the square of a number passed to it.
square = (x) -> x * x
If we call square 2, we'll get always get 4 back, regardless of when we call the function or whatever else might be happening in our program. Calling square doesn't affect (or even know about) any other part of our app — it only works with the number given to it. If we replaced every instance of square 2 in our code with the literal number 4, our program would run exactly the same.
Here's another example:
isStudent = (person) -> person.role == 'student'
Our isStudent function is pure. It only knows about the data passed to it. It doesn't have any side effects like changing the value of a variable or outputting something to the screen. It will always return the same value given the same person, like the one shown below:
person = name: 'Riley Blue' username: 'rileyblu' role: 'student'
Here's one last easy example of a pure function:
getNames = (people) -> people.map (person) -> person.name
Our getNames function is pure, for the same reasons square and isStudent are pure. This function doesn't change people, it only returns a new list built up from data in people. Array.map is pure, too.
Examples of Impure Functions
Let's jump over to PHP for four simple examples of impure functions.
function is_registered_for_classes($student) { global $registered_students; return in_array($student, $registered_students); }
The is_registered_for_classes function is impure because it relies on data not explicitly passed to it — in this case, the global variable $registered_students. If I saw a call to is_registered_for_classes somewhere in a program, I'd have no idea just from its name, inputs, or return value that it relied on a global value.
Worse still, if the state of $registered_students changes, we have no guarantee that is_registered_for_classes will return the same value. Impure functions like this are risky, because other unrelated parts of our program are in a position to hose us — more on that below.
function register_student($student) { ClassRoster::add($student); $student->has_registered_for_classes = true; }
The register_student function isn't pure, either. It's working with global state (although gussied up as a static object method, which makes some programmers feel better about their life). It's also assigning a new value to an existing object property, or mutating it, which is a textbook side effect.
Other parts of the program may rely on or behave differently based on the value of has_registered_for_classes, and assigning a new value to it may cause unexpected and unwanted (wait for it) side effects elsewhere in our app.
In functional programming, data is immutable. You can't change the value of something, but you can create new, remixed copies of data. I'll talk about immutability in my next post.
function course_enrollment_is_good() { return ClassRoster::getNumberOfEnrolledStudents() >= 25; }
This function is impure, because its result depends on the global state of ClassRoster. Calling course_enrollment_is_good at different times might give us a different result. We lose referential transparency: we couldn't go through our code and replace all instances of course_enrollment_is_good() with true, for example.
function say_hello() { echo 'Holler!'; }
This function is impure because it has the side-effect of printing text to the screen. Yes, input and output are side effects to be avoided and minimized.
Here's a tip: if you've written a function that takes no parameters or has no return values, you've written an impure function that depends on program state, has side effects, or both.
Avoid the void!
Here's three reasons why using pure functions can be good for your code:
They're easier to figure out and debug.
Realtalk — of all the things that aggravate the hell out of me as a programmer, the worst is action at a distance. Wikipedia's got a pretty decent definition:
Action at a distance is an anti-pattern (a recognized common error) in which behavior in one part of a program varies wildly based on difficult or impossible to identify operations in another part of the program.
Basically, action at a distance is what happens when one part of your program does something weird and screws up another part way over there because the two parts happen to both mess with a resource that has boundary issues. Global variables are a classic example of this and my least favorite thing in the whole world.
How many times have you called a function someone else wrote, only for it to do something (or require something) totally non-obvious from reading the function name and parameters? Me too.
Pure functions are straight up. With pure functions, what you see is all you get, all the time. Because they can only create transformed copies of data explicitly passed to them, you don't have to try and guess at global state or step through side effects in your head when working out what a pure function does.
If something goes wrong or the function returns jacked-up data, the haystack you have to sift through to find the problem is way smaller.
They're safer.
Because pure functions can't change program state or affect things outside of themselves, there's a much, much lower risk of a function call having unintended consequences on another part of your app. (This is doubly true in more "pure" functional programming languages like Clojure, where data is immutable: once a reference to a value is stored, that value can never change, and functions that use that value can always depend on it.)
Contrast this with a lot of procedural and object-oriented code, where functions often freely meddle with shared resources (global variables, static methods, object properties, and the like) on the other side of the room and don't have the courtesy to let anyone else know.
They're cacheable.
Because pure functions always return the same value given the same input, the results of calling those functions can be aggressively cached by the programmer or by the compiler. For expensive and time-consuming functions, being able to re-use the results of previous calls instead of calling the function again is a big deal.
This is called memoization.
What about state and input/output?
Alright, I know what you're going to say. You can't make literally every function pure. You can't always write a program where none of the functions do things like change program state, call out to other parts of the program, or produce side effects. So what do you do?
Well, there's two things you can do.
First, you can often rewrite your stateful, side effect-producing functions to be pure functions. This can be tough to do in mixed-paradigm languages that don't always make writing pure functions easy or idiomatic (like PHP), but a lot of the time you can.
Sometimes, you can improve things a lot just by explicitly passing dependencies to functions (such as the class roster), or by avoiding assignment statements. I'll talk more about specific refactoring strategies in a later post.
Second, functional programming languages isolate and minimize parts of the program that deal with state and input/output. In mixed-paradigm languages like Javascript or PHP, sometimes the best you can do is put that stuff in its own namespace and marking it with a big red sign saying "watch out!"
More "pure" functional programming languages, though, have more rigorous ways of safely dealing with state and side-effects, such as Clojure's software transactional memory and transient data structures.
How those things work is more than you need to know right now — just keep in mind that functional programming languages can be pretty strict about dealing with state and side effects.
Summary and Next Time
In this post, I introduced the concept of pure functions. Pure functions are functions that have no side effects like changing program state, don't work with data other than what's given to them, and always return the same result given the same input.
Like the other features of functional programming, pure functions make our code quicker to reason about, easier to debug, and safer to use.
Plus, because pure functions always return the same output when fed the same input, developers (and compilers) can cache the results of those function calls and bring down the overhead in our app.
Next time, we'll talk about my favorite part of functional programming: immutable data!


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Melting Pot @UCLPartners
In the fast-paced environment of health and care, it is very easy to feel disconnected from one another as we grapple with the challenges of increasing specialisation, constant reporting and inspections, and the seemingly relentless need to be seen to deliver.
Eating lunch together, and chatting over some common ground is a fantastic chance for us all to see things a little differently and help us make better connections, be better colleagues, and to better serve our patients and populations.
Melting Pot @UCLPartners is based upon the model developed by Kaleidoscope Health & Care and supported by the NHS London Leadership Academy.
Who can join in?
Lunches are open to anyone working in health, social care, voluntary sector and relevant academic organisations in or around the UCLPartners geography, or nationally.
What is it?
Each lunch is a 90-minute discussion forum with no other agenda, just bringing together diverse groups from across health and care, different disciplines, backgrounds, and settings. The aim is to discuss a particular topic of shared interest, and most importantly, enjoy lunch – all under the Chatham House rule.
Our guest speaker for the lunch is Paul Burstow, former Minister for Care Services, who will be helping us consider the question:
"Is maintaining a National Health Service too expensive, and too much of a sacrifice for the rest of the care system?"
Paul Burstow served for two and a half years as a Minister of State for Care Services in the Department of Health during the Coalition Government, where he had an extensive brief including care for the elderly, adult social care, mental health, cancer, diabetes and learning disability programmes. He was also responsible for reforming social care law, drafting the government's mental health strategy, the return of public health to local government, and the creation of health and wellbeing boards.
Please Note We need to share names and mail addresses of participants, so please make sure you are happy for others attending to know this before registering, and please be available promptly at 12:30pm and for the whole lunch.
Please email organiser (firstname.lastname@example.com) with any dietary requirements.


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Miley Cyrus Vs Taylor Swift Feud: Cara Delevingne Playing Peace Maker
Miley Cyrus and Taylor Swift aren't necessarily besties at the moment, but Cara Delevingne has plans to fix their on-going rift. An insider EXCLUSIVELY reveals to HollywoodLife.com just how the model plans on ending Miley and Taylor's drama once and for all!
Cara Delevingne, 23, to the rescue? The stunning supermodel is friends with Miley Cyrus, 22, and Taylor Swift, 25, so, it's only natural she knows about their latest drama. However, a source tells HollywoodLife.com EXCLUSIVELY that Taylor and Miley's fight is a 'nightmare' for the peace loving Brit and she plans on helping Miley and Taylor end their feud. To hear more about Miley Cyrus dissing Taylor Swift, head over to iTunes and download the latest HollywoodLife podcast for free!
Cara isn't one for drama and the supermodel isn't looking to participate in the latest feud between her two friends. "Cara is tight with Miley and Taylor, so this is a nightmare for her," a source tells HollywoodLife.com EXCLUSIVELY. "She hates when people don't get along. She's trying to get them to talk and work it out." The 23-year-old is doing her best to stop the fight between Miley and Taylor, but she might have her work cut out for her. Miley's comment about Taylor's violent "Bad Blood" music video in her Marie Claire spread isn't the first time she threw shade at the popstar.
The 22-year-old left a bad taste in Taylor's mouth when she posted a silly lesbian face-swap picture of Tay and Justin Bieber, 21, on Instagram. So, it's safe to assume the two may not even want to talk to each other. However, Cara has the perfect plan if they won't talk it out. "If they won't talk she's threatening to trick them into hanging out," our source continued to say. "She wants them to be friends and get over all the drama. And she might get her way, she's very convincing." Hopefully Cara works her magic before Miley and Taylor face off at the VMA's.
Taylor Swift Thinks Miley Cyrus Is Jealous Of Her Success
So far, Taylor hasn't publicly responded to Miley's diss towards "Bad Blood." But, a source told HollywoodLife.com EXCLUSIVELY that the 25-year-old thinks Miley is "desperate for attention and will do anything to get it. She wants the spotlight on her and taking shots at Taylor is a good way to do it." Makes sense — after all it was only a few months ago that Miley was pointing a gun at Zoe Kravitz in a music video. Either way, we do hope Cara can fix this feud.
But what do you think HollywoodLifers? Should Cara intervene in Miley and Taylor's feud? Let us know!
— Allison Swan


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- Automotive
- Black banks
- black business
- Black Economics
- Black Food
- black owned
- Black Owned Vegan Businesses
- BlackOutMonth2020
- Body Butters
- California Black Businesses
- Chicago
- Cosmetics
- Farmers
- Feminine Care
- Gas Stations
- Gun Ranges
- Gun Stores
- Hygiene
- JuneteenthBlackout
- JuneteethBlackOut
- los angeles
- Make-up
- Men's Grooming Business
- Oral Care
- Restaurants
- Shaving Cremes
- Skincare Lines
- support black businesses
- Toilet Paper
- Vegan Business
- Women's Products
What Are You Selling? (Hint: It's Not Your Blog)
What Are You Selling? (Hint: It's Not Your Blog)
I get a lot of emails from ladies who are in the beginning stages of starting a side hustle that fits their passion and aligns with their ideal life. In fact, over half of them say something like:
I love what you're doing on Happy Black Woman, how can I do that, too?
The problem is, most people mistakenly think that this blog IS my business, when it is certainly NOT. I love writing here to document my own journey and encourage other women along in their path. That's my main motivation for maintaining the site. What keeps it a priority on my list to be here every day is that I have also been able to earn a little bit of money from helping people through my work here.
This Blog is Not a Business
The blog itself? Nah, it's not a business. And neither is yours.
I have a few advertisements and affiliate links on this site, but the revenue is hit or miss. Unreliable, but nice when a check comes in. My real business is writing, speaking, coaching, training, consulting. That has never changed. I get paid for my writing when I sell books and ebooks. I get paid when nonprofits and associations hire me to come speak at their conferences. I get paid when people book a coaching session with me to get clarity around their career, business or personal growth. I get paid to conduct workshops and webinars on leadership, career development, and social media. I get paid to help professionals build their personal brands for career advancement. I get paid to help entrepreneurs and small business owners develop their online marketing strategy.
Those are the products and services that I'm selling. My main site (along with this blog) allows me promote myself as someone who adds value to people's lives through information and inspiration. In turn, that leads to more clients and customers. Same thing with my Twitter accounts and Facebook pages. They all serve a purpose in helping me sell myself so I can help more people and fulfill my personal mission.
How This Applies to You
If you want to build a successful business on the side – or turn your side hustle into your full-time hustle, you have to be very clear on what you're selling. You want to create a website like mine? Fine. (Here's my free ebook on how to start a blog.) Write to help people. Write to share your story and inspire others. Just remember that you cannot sell motivation alone. If you're using it to promote your business, then your blog has to be a vehicle for something that actually brings in money.
Figure out what you're selling. (Here's a post I wrote on how to define your side hustle.) Then use your blog to promote your products or services to people who might be interested in buying.
Does this make sense to you? Is there anything I can clarify further about this process?
You may also like:
The post What Are You Selling? (Hint: It's Not Your Blog) appeared first on Happy Black Woman.


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Vitamin C (l-ascorbic acid) is a potent antioxidant and cellular reductant present at millimolar concentrations in plants. This small molecule has roles in the reduction of prosthetic metal ions, cell wall expansion, cell division, and in the detoxification of reactive oxygen generated by photosynthesis and adverse environmental conditions. However, unlike in animals, the biosynthesis of ascorbic acid (AsA) in plants is only beginning to be unraveled. The previously described AsA-deficient Arabidopsis mutant vtc1 (vitamin c-1) was recently shown to have a defect in GDP-mannose pyrophosphorylase, providing strong evidence for the recently proposed role of GDP-mannose in AsA biosynthesis. To genetically define other AsA biosynthetic loci, we have used a novel AsA assay to isolate four vtc mutants that define three additional VTC loci. We have also isolated a second mutant allele of VTC1. The four loci represented by the vtc mutant collection have been genetically characterized and mapped onto the Arabidopsis genome. The vtc mutants have differing ozone sensitivities. In addition, two of the mutants, vtc2-1 and vtc2-2, have unusually low levels of AsA in the leaf tissue of mature plants.
MUTANTS have been extremely valuable tools for the dissection of biosynthetic pathways ever since the pioneering work on arginine-requiring mutants of Neurospora crassa in the 1940s by Srb and Horowitz. In plants, mutants have greatly facilitated the elucidation of a wide variety of complex pathways, including (for example) the biosynthesis of tryptophan (Radwanski and Last 1995), the hormone abscisic acid (Koornneefet al. 1998), carotenoids (Buckner and Robertson 1993; Norriset al. 1995), and fatty acids (Miquel and Browse 1998). Although mutants have been utilized to uncover a number of plant primary and secondary metabolic pathways, many more pathways have yet to be unraveled.
The antioxidant and cellular reductant vitamin C (l-ascorbic acid; AsA) has diverse physiological roles in plants (for a review see Smirnoff 1996). Despite the importance of this abundant small molecule in plant physiology and animal health, only very recently has significant progress been made toward elucidation of the plant AsA biosynthetic pathway (Wheeleret al. 1998). In contrast, the AsA biosynthetic pathway in animals has been known for some time. Animals generate AsA via the intermediates d-glucose, d-glucuronic acid, l-gulonic acid, and finally l-gulono-1,4-lactone, which is then oxidized to AsA. In this pathway the stereochemistry of the carbon skeleton of the primary substrate glucose is inverted in the final product (for a review see Burns 1967). A similar pathway was proposed for plants (Isherwoodet al. 1954). However, it was later shown that inversion of the glucose carbon skeleton does not occur during plant AsA biosynthesis (Loewus 1963). A noninversion pathway with the intermediates d-glucosone and l-sorbosone was then proposed to accommodate the observed stereochemistry (Loewus 1988) although there has been no recent evidence for this pathway. Strong biochemical evidence for a novel plant AsA biosynthetic pathway was recently published. This proposed pathway, with conversion of d-mannose to l-galactose via a little known GDP-mannose-3,5-epimerase as a key step, does not invoke inversion of the carbon skeleton of glucose (Wheeleret al. 1998). Furthermore, the Smirnoff/Wheeler pathway leads to conversion of l-galactose to l-galactono-1,4-lactone, the substrate for galactono-1,4-lactone dehydrogenase (GLDH) enzyme (Ôbaet al. 1995; Østergaardet al. 1997), which oxidizes l-galactono-1,4-lactone to AsA.
We are using Arabidopsis thaliana mutants that are deficient in AsA to identify genes involved in the maintenance of AsA levels in plants. The identification of such genes will facilitate critical analysis of the plant AsA biosynthetic pathway as well as identify the pathway(s) of AsA catabolism. Screens for ozone-sensitive A. thaliana mutants led to the isolation of many mutants, two of which contain only ~25–30% of wild-type (wt) AsA (Conklinet al. 1996; P. L. Conklin and R. L. Last, unpublished data). Complementation analysis that relied upon the ozone-sensitive phenotype indicated that these lines have nonallelic recessive mutations and they were named vtc1-1 and vtc2-1 (vtc; vitamin c). The vtc1-1 mutant has a reduced ability to convert d-glucose (the primary substrate for AsA) into AsA, suggesting that this locus encodes a biosynthetic pathway enzyme (Conklinet al. 1997). Indeed, the VTC1 locus was recently cloned and shown to encode a GDP-mannose pyrophosphorylase (Conklinet al. 1999). This enzyme catalyzes the conversion of d-mannose-1-P to GDP-mannose, a step in the Smirnoff/Wheeler AsA biosynthetic pathway (Wheeleret al. 1998). VTC1 is the first locus that has been genetically identified as having a role in plant AsA biosynthesis, and our results provide strong support for the role of GDP-mannose in AsA biosynthesis.
As the recovery of AsA-deficient mutants from our screen for ozone-sensitive mutants was very low (2/100,000 M2 plants screened or ~0.002%), we turned to a more direct screen to identify additional VTC loci. As detailed below, a nitroblue tetrazolium (NBT)-based assay was developed and used to screen ethyl methanesulfonate (EMS)-mutagenized Arabidopsis seedlings for an AsA-deficient phenotype. The characterization of five new vtc mutants isolated by this screen is described below. Together, our collection of mutants represent four different VTC loci.
Plants and growth conditions: All wt and mutant A. thaliana lines used in this study are derived from the Columbia (Col-0) ecotype. Mutant lines used for both the quantitative measure of AsA and determination of ozone sensitivity were derived from at least one backcross to Col-0 wt. The vtc1-1 line was derived from four backcrosses while the vtc1-2 and vtc4-1 lines were each backcrossed twice.
Specific growth conditions varied depending on the experiment. Seeds of mutagenized M2 and F2 polymorphic mapping populations, as well as F2 seed used for segregation and linkage analyses, were plated on sterile nutrient medium without sucrose (PN; Haughn and Somerville 1986) and grown on a 24-hr photoperiod as previously described (Liet al. 1995). The F2 vtc/vtc mapping individuals were then transplanted to soil-less Cornell mix and grown under a 24-hr photoperiod with other conditions as described (Conklinet al. 1999). All plant material used for the quantitative measure of AsA were grown under a 16-hr photoperiod (Conklinet al. 1999). Mutant and wt plants to be fumigated with ozone were grown in a constant environment chamber under a 16-hr photoperiod as described (Conklinet al. 1996).
l-ascorbic acid assays: A qualitative AsA assay was developed that utilizes NBT as a reagent for the visual detection of AsA. Arabidopsis leaves ~3–8 mm in length are excised and laid on a sheet of chromatography paper. Whatman 3030-6185 paper (Whatman Ltd., Kent, UK) works well for this assay while generic brands do not. Each leaf is then squashed onto the chromatography paper using a curved metal weigh spatula. Ten microliters of a 1 mg/ml aqueous solution of NBT (Sigma, St. Louis) is then pipetted directly onto each squashed leaf. Within ~5 min, a bluish-purple formazan precipitate is visualized around each wt leaf. As the formazan tends to bleed through the chromatography paper, this precipitate can often be visualized better on the back side of the paper.
Two different quantitative AsA assays were used in this study: a previously described ascorbate oxidase-based spectrophotometric assay (Conklinet al. 1996) and an HPLC-based assay modified from the procedure of Kutnink et al. (1987) and Mitton and Trevithick (1994). For the detection of total AsA by an electrochemical (EC) assay coupled to HPLC, frozen samples were extracted using Tenbroeck tissue grinders (Lawson Mardon Wheaton Inc., Millville, NJ) in cold 5% metaphosphoric acid containing 0.54 mm Na2EDTA and samples were microcentrifuged for 10 min to remove solids. The lysate was diluted to 0.2% metaphosphoric acid, 0.54 mm Na2EDTA, 1 mm cysteine, and 1 mg/ml dithiothreitol (DTT); incubated for 60 min at room temperature prior to injection; and then microfuged for 5 min. Samples of the supernatant were injected onto a C-18 ODS column (BAS, W. Lafayette, IN) with a 0.02 m NaAcetate, 0.54 mm Na2EDTA, 1.5 mm N-octylamine mobile phase, and AsA was detected with a BAS LC-4B elecrochemical detector containing a glassy carbon working electrode. The applied potential was +0.5 V (oxidative) with the sensitivity set at 50 nA. The run time for each sample was 10 min with the AsA peak eluting at ~2 min and a wide DTT peak appearing at ~6 min.
Mutant screen: Col-0 wt seed was mutagenized with EMS as described (Barczaket al. 1995). M2 seed was suspended in 0.1% agar and hand-plated in a grid pattern on PN medium in 100-mm-diameter petri plates at a density of 100 seeds/plate. A single true leaf from each 2-wk-old plant was assayed for AsA using the NBT-based assay. The plants that showed a reduced formazan halo were then transplanted to soil and allowed to self-pollinate. Two-week-old M3 plants were retested for reduced AsA by the NBT-based assay as well as the ascorbate oxidase- and EC-HPLC-based assays. Mutants that contained ≤50% of wt AsA were subjected to further analysis.
Genetic analyses: To test whether the vtc mutation segregated as a single monogenic trait, F1 seed was obtained by pollination of VTC/VTC stigmas with vtc/vtc pollen or vice versa. F1 progeny were allowed to self-pollinate to obtain segregating F2 populations. Two-week-old plants from these populations were then scored using the NBT-based assay. To test for allelism, an F2 segregating population was obtained from a cross between two independently isolated ascorbic acid-deficient lines. Two-week-old F2 plants were then scored for AsA using the NBT-based assay. Two independently isolated vtc mutants were judged as nonallelic if F2 progeny with wt levels of AsA were obtained.
Each of the VTC loci was mapped onto the Arabidopsis genome by scoring genetic markers throughout the genome on vtc/vtc individuals (scored as Nbt−) from a polymorphic F2 mapping population generated by a cross between the VTC/VTC (Ler ecotype) and vtc/vtc (Col-0 background). Both microsatellite (Bell and Ecker 1994) and cleaved amplified polymorphic sequences (Konieczny and Ausubel 1993) were used as markers. VTC2, VTC3, and VTC4 were mapped using 50 vtc2-1/vtc2-1, 54 vtc3-1/vtc3-1, and 31 vtc4-1/vtc4-1 F2 individuals. Genetic map locations were calculated using the Kosambi mapping function (Koornneef and Stam 1992).
Assessment of ozone sensitivity: Ozone fumigations were performed on 2-wk-old vtc mutant and wt plants grown on a 16-hr photoperiod from 6:00 AM to 10:00 PM as previously described (Conklin and Last 1995). Plants were exposed to 400 ppb ozone for 8 hr from 9:00 AM to 5:00 PM. Photographs were taken 16 hr after the end of the exposure.
As described above, the AsA-deficient Arabidopsis mutants vtc1-1 and vtc2-1 were isolated by virtue of their ozone sensitivity and are part of a large collection of ozone-sensitive mutants. However as vtc mutants represented <5% of the ozone-sensitive mutant class, it was necessary to develop a direct AsA assay to obtain additional vtc mutants. We developed a simple and rapid screen for AsA-deficient Arabidopsis mutants based upon the ability of AsA to convert NBT to formazan. Five additional vtc mutants including new alleles of VTC1 and VTC2 as well as mutants that define two new complementation groups were identified with this technique.
A novel l-ascorbic acid detection method: Our new AsA assay utilizes the electron transfer dye NBT, which can be reduced by four electrons to yield the dark bluishpurple insoluble formazan. Purified AsA reduces NBT to the formazan, and the high AsA content in plant tissue has allowed us to take advantage of this property. In this method, Arabidopsis leaves are excised and then squashed onto chromatography paper causing the aqueous contents of the leaf to be wicked into the paper. Each squashed leaf is then treated with NBT, and the formazan precipitate is visualized around the (wt) leaf after ~5 min.
This simple assay was titrated to reliably differentiate between the AsA level in leaves from 2-wk-old wt and vtc1-1 plants as shown in Figure 1. Wt Col-0 Arabidopsis at 2 wk of age contains 3–4 μmol AsA/g fresh weight (FWT) while the AsA-deficient mutant vtc1-1 contains ~0.8–1.0 μmol AsA/g FWT (Conklinet al. 1996). Therefore this assay permits discrimination between the ~7 nmol of AsA in a single 2-mg wt leaf and the ~2 nmol of AsA in a single vtc1-1 leaf.
The efficacy of the NBT-based assay was tested by demonstrating that the presence of formazan (Nbt+) cosegregates with wt levels of AsA while the absence of formazan (Nbt−) correlates with low AsA levels in a population segregating for the vtc2-2 mutant allele. Individual F2 progeny from a population segregating for the vtc2-2 mutant allele (described below) were scored for NBT. Ten Nbt− progeny and 20 Nbt+ progeny were then assayed for total AsA using an EC-HPLC-based method. The Nbt− individuals were all AsA deficient while the Nbt+ individuals all contained higher levels of AsA (Figure 2). This demonstrates that vtc2/vtc2 mutant individuals can be accurately detected in a background of VTC2/VTC2.
Isolation of vtc mutants: Using this NBT-based assay, ~6300 M2 A. thaliana plants (ecotype Col-0) from an EMS-mutagenized M2 population were screened for an AsA deficiency. Forty-eight of the M2 individuals scored as negative in this primary screen. For each putative mutant line, M3 generation populations were rescreened for an AsA deficiency using the NBT-based assay, an EC-HPLC-based assay (Kutnick et al. 1987; Mitton and Trevithick 1994), and a spectrophotometric assay (Conklinet al. 1996). Five of the original 48 putative mutant lines rescored as Nbt− in the M3 generation. These 5 lines were independently isolated from different pools of mutagenized seed, and all exhibited AsA deficiencies (based on the spectrophotometric and EC-HPLC assays). Therefore, 5/6300 M2 plants (~0.1%) screened using the NBT-based assay were AsA deficient as compared to a ~0.002% recovery of vtc mutants with the ozone sensitivity-based screen. The fact that all mutants rescoring as Nbt− in the M3 generation were AsA deficient underscores the specificity of the NBT-based screen for AsA mutant identification.
Genetic analysis: In addition to using the NBT-based assay to identify new mutants, it was also used for analyses of genetic segregation and allelism. In both cases, individual progeny from two independent crosses per mutant line were scored for the presence (Nbt+) or absence (Nbt−) of wt levels of AsA.
Our data indicate that the AsA deficiency in the mutants vtc1-2, vtc2-1, vtc2-2, vtc3-1, and vtc4-1 is conferred by single monogenic recessive traits. As shown in Table 1, F2 progeny from crosses between three of the vtc mutant lines (vtc1-2, vtc2-2, vtc3-1) and wt Col-0 segregate in a statistically significant 3:1 ratio of Nbt+:Nbt− plants (P ≥ 0.2). In contrast, the F2 progeny from the cross between Col-0 wt and vtc2-1 yielded an unexpectedly high number of Nbt+ individuals (P = 0.003) while the F2 progeny of the cross between Col-0 and vtc4-1 included a somewhat high number of Nbt− individuals (P ≤ 0.05; Table 1). These data are unlikely to result from a gene dosage effect as both VTC2/vtc2-1 and VTC4/vtc4-1 heterozygotes contain wt levels of AsA (data not shown). The reason(s) for the observed anomalies in these segregation ratios is not clear. However, crossing both these mutant alleles to a different wt ecotype (Ler) yielded F2 progeny in the expected 3:1 ratio of Nbt+:Nbt− suggesting that the AsA deficiencies in these mutants are indeed conferred by single monogenic recessive traits.
The phenotypes of F2 progeny from crosses between the mutant vtc2-3 and Col-0 were somewhat skewed toward the presence of Nbt− individuals. To test the hypothesis that this is a gene dosage effect, AsA levels were quantitatively measured in two sets of pooled F1 progeny from the cross (vtc2-3 × Col-0). As seen in Figure 3, these F1 heterozygotes contain levels of AsA intermediate between the two parents, suggestive of a gene dosage effect. Given the fact that the NBT-based assay is only semi-quantitative, some of the VTC2/vtc2-3 F2 progeny were probably scored as Nbt− resulting in the observed skewed ratio of Nbt+/Nbt− individuals.
We tested for allelism between the AsA-deficient Arabidopsis mutants and these results indicate that the vtc mutants represent four different loci: VTC1, VTC2, VTC3, and VTC4. As shown in Table 2, both wt (Nbt+) and mutant (Nbt−) individuals were found in the segregating F2 progeny from crosses between nonallelic mutants such as vtc1-1 and vtc2-2. In contrast, the F2 segregating progeny from a cross between mutants harboring mutations at the same locus scored as mutant. A compilation of the segregation data in Table 2 shows that there are two vtc1 mutants and three vtc2 mutants, as well as single vtc3 and vtc4 alleles.
Genetic map positions: The F2 segregation data were extended by genetically mapping VTC2 through VTC4 (Figure 4). The loci were mapped using polymorphic F2 mapping populations generated from crosses between these mutants and the wt line Ler, which contains well-characterized microsatellite polymorphisms (Bell and Ecker 1994) and cleaved amplified polymorphic sequences (CAPS; Konieczny and Ausubel 1993) compared with Col-0. The vtc2-1 mutation was found to map to a position on chromosome 4 ~3 cM centromere distal to CAPS marker WU95, which resides at position 71.70 on the latest Arabidopsis recombinant inbred (RI) genetic map (http://genome-www.stanford.edu/Arabidopsis/ww/Aug98RImaps/index.html) and ~5 cM centromere proximal to CAPS marker PRHA (position 76.17). vtc2-2 and vtc2-3 map to the same region as vtc2-1 (S. R. Norris and R. L. Last, unpublished results). VTC3 was mapped to a position on chromosome 2 close to VTC1, ~4 cM centromere distal from microsatellite marker nga168 (position 73.01). The VTC4 locus was mapped to the top of chromosome 4 ~2 cM centromere distal from microsatellite marker nga172 (position 6.83). VTC1 was previously mapped on chromosome 2 to a position 0.9 cM centromere distal from CAPS marker m429 (Conklin et al. 1996, 1999).
l-Ascorbic acid levels in vtc mutants: The new vtc mutants described above were identified as having a diminished ability to reduce NBT to formazan. To quantitatively measure the AsA deficiencies in these mutants, a spectrophotometric method was used to measure total AsA in 2-wk-old rosettes from each of the vtc mutant lines. The lines used in this analysis have all been backcrossed at least once to the wt progenitor to segregate away unlinked mutations. Our results indicate that the vtc mutants contain 1/3 to ½ the total AsA present in the wt Col-0 progenitor as shown in Figure 5.
In plants, AsA levels are known to increase upon transition from the vegetative to reproductive state (Gander 1982). To determine whether such an increase occurs in the wt and vtc mutants, total AsA was measured in mature rosette leaves, immature green siliques (seed pods), and the inflorescence (containing a mixture of opened and unopened flowers) of 6-wk-old plants. As shown in Figure 6, reproductive tissues from wt (green siliques and inflorescences) contain approximately twice the amount of AsA found in wt mature leaves. All the vtc mutants are also able to maintain higher levels of AsA in the reproductive tissues relative to that in leaves. In fact, the floral tissues of these "AsA-deficient" mutants contain ≥3 μmol/g FWT AsA, matching the levels found in rosette leaves of wt.
An interesting anomaly was uncovered upon comparison of the AsA levels in the leaves from 6-wk-old wt and vtc mutants. In mature (fully expanded) leaves, the majority of vtc mutants maintain AsA levels at ~40% (~1.7 μmol/g FWT) of wt. vtc2-1 and vtc2-2 represent an exception and mature leaves from these two mutant lines have unusually low levels of AsA (~10% of wt; ~0.40 μmol/g FWT). vtc2-1 is also severely AsA deficient in younger leaves and cauline (stem) leaves from older plants. In summary, 6-wk-old vtc2-1 and vtc2-2 have a very severe AsA deficiency in leaves while siliques and inflorescences from these same plants as well as leaves from 2-wk-old plants are not as severely deficient. This suggests either that there is an underlying difference(s) in AsA metabolism in these different tissue types or that VTC2 is a regulatory gene. As described below, the vtc mutants also have unexpected differences in their ozone sensitivities.
Ozone sensitivity: The anthropogenic air pollutant ozone (O3) is a well-documented cause of oxidative stress in plants. Ozone enters the plant through open stomata and then presumably degrades into ·O−2, H2O2, and OH· in the aqueous apoplastic environment (Heath 1994). The wt Col-0 ecotype of Arabidopsis is quite tolerant to O3, probably because these plants mount an effective antioxidant response (Sharma and Davis 1994; Conklin and Last 1995). In contrast, the AsA-deficient mutant vtc1-1 is extremely O3 sensitive with visible injury including lesion formation, enhanced chlorosis, and/or tissue collapse (Conklinet al. 1996). Severely injured leaves do not recover after the exposure; however, immature leaves emerging during the treatment are not visually injured, presumably due to a lack of fully functioning mature stomata (Conklinet al. 1996).
To test the hypothesis that AsA is important for protection against O3 injury, we examined the sensitivity of the vtc mutants and found a surprisingly wide range of response to this source of oxidative stress. Two-week-old vtc and wt plants were exposed to 400 ppb O3 for 8 hr. Directly before this treatment, one set of plants was moved to a control chamber with very similar environmental conditions but where O3 was depleted by activated charcoal filtration. Photographs were taken of representative treated and control plants 16 hr after the end of the O3 exposure (Figure 7) and tissue from the control plants was assayed for total AsA (Figure 5). Surprisingly, the different AsA-deficient mutant lines have varied O3 sensitivities, sometimes even within an allelic series. As previously reported, the vtc1-1 mutant is very sensitive to O3 damage (Conklinet al. 1996). Consistent with our observation that vtc1-2 has the same missense mutation as vtc1-1 (Conklinet al. 1999) this mutant exhibits a similar sensitivity. This injury is seen as total collapse and death of both cotyledons and fully expanded leaves. The different vtc2 mutants have highly varied O3-sensitive phenotypes. vtc2-1 is as sensitive to O3 as the vtc1 mutants, and vtc2-3 appears to be somewhat sensitive as O3 exposure of this mutant leads to partial collapse of at least one fully expanded leaf per plant. Curiously, vtc2-2 is not visibly injured by this high dose of O3 despite that fact that it contains a steady state level of AsA very similar to vtc2-1 (Figure 5). As with vtc2-3, the mutants vtc3-1 and vtc4-1 are only slightly more O3 sensitive than the wt.
A rapid and simple assay has been developed to semi-quantitatively measure AsA in leaf tissue using NBT, which is reduced by AsA to generate a purple formazan precipitate. We have defined assay conditions whereby formazan is formed when wt Arabidopsis leaves are treated with NBT, but little or no formazan is visible when assaying the AsA-deficient mutant vtc1-1. This assay was utilized to screen populations of EMS-mutagenized 2-wk-old seedlings. Five independent mutant lines were isolated that had a heritable Nbt− phenotype. It was somewhat surprising that these five Nbt− lines were all AsA deficient as other cellular constituents (such as superoxide) are also able to reduce NBT to formazan. However, as ascorbic acid is present in very high concentrations relative to other possible NBT-reducing agents and the assay was titrated using vtc1-1 and wt plants, the screen was most likely biased toward the isolation of additional vtc mutants. The five new vtc mutants represent four VTC loci. These are in addition to two vtc mutants previously isolated as O3 sensitive. In total, there are two vtc1 mutant alleles, three vtc2 mutants, as well as vtc3-1 and vtc4-1. All four loci were genetically mapped, with VTC1 and VTC3 found to be linked at the bottom of Chr 2, VTC2 on Chr 4, and VTC4 at the top of Chr 3.
The vtc mutants are valuable tools for studying the regulation of AsA accumulation in plants, including elucidation of the biosynthetic pathway. Identification of AsA-deficient Arabidopsis mutants that harbor mutations in AsA biosynthetic enzymes would lend strong genetic evidence for the pathway proposed by Smirnoff and colleagues (Figure 8; Wheeleret al. 1998) or an alternative pathway. We have recently shown that VTC1 encodes GDP-mannose pyrophosphorylase, a predicted enzyme in the Smirnoff/Wheeler pathway (Conklinet al. 1999). Preliminary labeling data suggest that the vtc2 and vtc3 mutants have reduced ability to convert mannose to AsA (S. R. Norris, G. L. Wheeler and N. Smirnoff, unpublished data).
Isolation and analysis of the AsA biosynthetic genes and gene products should also help to clarify the subcellular locale(s) of the proposed pathway. The enzymes responsible for the generation of fructose-6-P are well-studied glycolytic enzymes. In plants, glycolysis occurs in the cytosol and at least the first half of the glycolytic pathway is also present in the plastid. Isoenzymes encoded by different nuclear genes are thought to be responsible for catalysis of each of the steps in the two parallel pathways (for a review see Plaxton 1996). In yeast, phosphomannose isomerase, phosphomannomutase, and GDP-mannose pyrophosphorylase are all localized to the cytosol (Kepes and Schekman 1988; Smithet al. 1992; Hashimotoet al. 1997). Similarly, the Arabidopsis GDP-mannose pyrophosphorylase protein inferred from the VTC1 coding sequence contains no obvious targeting sequences, suggestive of a cytosolic localization of this enzyme in plants (Conklinet al. 1999). In contrast, the last biosynthetic enzyme in the pathway, GLDH, is found in the mitochondria of plants (Ôbaet al. 1995; Østergaardet al. 1997). It is plausible that portions of the plant AsA biosynthetic pathway could occur in at least three different subcellular compartments, the plastid, cytosol, and mitochondrion.
In addition to aiding in the elucidation of the AsA biosynthetic pathway and its cellular biology, the vtc mutants should help us to better understand the physiological roles of AsA. We have examined the AsA level in both mutant and wt plants at different ages and in different tissue types and have measured the O3 sensitivity of the different vtc mutants. At 2 wk of age, all of the vtc mutants contain ≥33% of wt AsA. As AsA has many roles, both as an antioxidant and as a cellular reductant, it is plausible that more severe decreases in AsA would be lethal. Consistent with this hypothesis, a nonsense mutation at the VTC1 locus, which is predicted to abolish the activity of the GDP-mannose pyrophosphorylase, was identified as a cytokinesis-defective embryo lethal (cyt1-2; Nickle and Meinke 1998; Somervilleet al. 1998; W. Lukowitz and C. Somerville, personal communication). While this is suggestive of an essential requirement for AsA, the GDP-mannose synthesized by GDP-mannose pyrophosphorylase (Figure 8) is used in protein glycosylation and cell wall synthesis in addition to AsA synthesis. Because of the predicted pleiotrophy of a mutant at this step in the pathway, the embryo-lethal phenotype of cyt1-2 is presumably due to a combination of factors including defective protein glycosylation and cell wall structure as well as a lack of AsA.
As mentioned above, the vtc mutants are all only partially AsA deficient. These deficiencies could result from mutations causing partial inactivation of wt functions (leaky mutations) and/or functional redundancy of some of the genes involved in AsA biosynthesis. For example, we believe that the mutant vtc1-1 is partially deficient due to a leaky mutation. vtc1-1 contains a missense mutation that is predicted to alter a highly conserved proline residue in the GDP-mannose pyrophosphorylase amino acid sequence. As this mutant still contains ~33% of wt AsA and partial GDP-mannose pyrophosphorylase activity, we hypothesized that the nonlethal vtc1-1 mutation negatively affects but does not abolish this enzyme activity (Conklinet al. 1999). In 6-wk-old wt Arabidopsis, the AsA levels in reproductive tissues (open and closed flowers and green siliques) are approximately double the amount assayed in mature leaves. It is well established that AsA levels are generally high in plant reproductive organs relative to vegetative tissue (reviewed in Gander 1982). This is likely due to a greater need for AsA in these tissues. Reproductive tissues have higher metabolic rates, which would necessitate the need for increased levels of antioxidants. In addition, such tissues have higher rates of cell expansion and division and AsA is thought to have a role in these processes (for a review see Smirnoff 1996). While the control mechanism(s) orchestrating this organ-specific up-regulation is not known, it has been shown that carbohydrate status has a role in governing AsA levels in plants (Smirnoff and Pallanca 1996). The relative higher level of AsA in flowers and siliques over that in mature leaves is maintained in the vtc mutants despite their overall AsA deficiency, implying that the mutants retain the control mechanism(s).
vtc2-1 and vtc2-2 rosette and cauline leaves from 6-wk-old plants contain ~10% of wt amounts of AsA while the amount of AsA in the seedlings, inflorescence, and green siliques of these mutants is similar to that of vtc1-1 and vtc1-2 (~33% of wt). Despite the very low leaf levels of AsA in the leaves of these older plants, this tissue looks quite normal, suggesting that other antioxidants are compensating in this tissue or that high levels of AsA are not necessary in the leaves of older plants. We have preliminary evidence that vtc2-1 is defective in AsA biosynthesis (S. R. Norris, G. L. Wheeler and N. Smirnoff, unpublished data). If VTC2 encodes a biosynthetic enzyme, one could imagine specific protein alterations that would affect enzymatic activity as flux through the pathway changes. For example, a mutation increasing the Km of an AsA biosynthetic enzyme might cause a severe decrease in AsA only in tissues where carbohydrate is limiting. Limited carbon may be available for AsA synthesis in leaves of older plants (i.e., it is getting shunted to the reproductive tissues) while carbon may not be limiting in young leaves, growing siliques, and the inflorescence. As vtc2-1 and vtc2-2 have low AsA in the leaves of older plants, but not in these other tissues, we hypothesize that these two mutant alleles might represent such a biochemical defect. The vtc2-3 mutant does not follow this pattern as the AsA level in mature leaves of this mutant are not severely affected (~33% of wt). An alternative hypothesis is that VTC2 is a regulatory locus rather than encoding a biosynthetic enzyme. Given the fact that AsA levels in wt vary in different tissues, specific mutations at a site involved in this regulation could adversely affect AsA levels in a tissue-specific manner. The VTC2 locus is currently being cloned; identification of the VTC2 gene product and the nature of the vtc2 mutations will hopefully shed some light on this interesting question.
In addition to the differing AsA phenotype of the vtc mutants, we have also found unexpected differences in O3 sensitivity between the mutants. O3 generates oxygen free radicals within plant tissue and the mutants vtc1-1, vtc1-2, and vtc2-1 are all quite O3 sensitive relative to wt. In contrast, the other vtc mutants are also AsA deficient, while exhibiting only mild O3 sensitivity. Given that the mutants vtc2-3, vtc3-1, and vtc4-1 have slightly higher levels of AsA (~50% of wt) than the more O3-sensitive vtc1-1, vtc1-2, and vtc2-1, it is possible that the threshold between O3 resistance and sensitivity is just below this amount. Alternatively, side products and/or metabolic intermediates with antioxidant activity could be accumulating in vtc3-1 and vtc4-1, affording some protection against the free radicals generated by O3. The difference in O3 sensitivity of the mutants vtc2-1 and vtc2-2 cannot be explained by a threshold mechanism as they have very similar levels of AsA yet vastly different sensitivities to O3. The physiological basis for this difference remains to be determined.
Identification of a broad range of mutants has revealed the complexity of AsA physiology and metabolism in plants. As several of the vtc mutant alleles have a response to O3 similar to that of wt, they would not have been isolated in a screen for O3-sensitive mutants. Alternatively, isolation of several vtc mutants by a direct screen has demonstrated the utility of the NBT-based assay as a facile method for the specific isolation of AsA-deficient mutants. Identification of VTC1 as encoding GDP-mannose pyrophosphorylase provided strong genetic support for the proposed role of GDP-mannose in AsA biosynthesis. Likewise, cloning of the VTC2, VTC3, and VTC4 loci should also add greatly to our understanding of how plants synthesize AsA.
We thank Richa Agarwal for her technical assistance in the mapping of VTC4. This work was supported by grant 96-35100-3212 from the Plant Responses to the Environment Program of the U.S. Department of Agriculture National Research Initiative Competitive Grants Program to R.L.L. and P.L.C. and by National Institutes of Health postdoctoral fellowship GM18945-01 to S.R.N.
Communicating editor: K. J. Newton
- Received February 12, 1999.
- Accepted October 13, 1999.
- Copyright © 2000 by the Genetics Society of America


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Friday, May 30, 2008
Me... While I'm very proud of him, and I understand it's great to be validated.... I'm sorta thinking... But I thought the season was almost over???
6 comments:
- 3boys247 said...
How exciting! Congratulations TMAC!
- May 31, 2008 at 6:01 AM
Woo hoo!! Congrats!! Our youngest son just made All-Stars too!! Our oldest son play competitive year round select baseball so we are so excited that little Adam can have his own special time now. I need to create a special post for him because this is a big deal. Go All-Stars!!
~Jill :)
- May 31, 2008 at 6:16 AM
- Hipmomofboyz said...
how exciting for your hubby and son...
You know I love baseball, but when Kyle's team got knocked out of the playoffs Thursday night. let's just say I did not shed any tears....lol
- May 31, 2008 at 2:27 PM
- Kristen said...
YEA and OH NO!! All at the same time!
Are you finding yourself craving concession stand hot dogs yet?
- May 31, 2008 at 4:21 PM
- The Joys of having Boys said...
Congrats to dad!! You will have to tell me how you do it. J#1 is having his first game this week,and I only have one in a sport at this time. It feels like I am running in circles.
- June 1, 2008 at 2:56 PM
- jennwa said...
That is great. Congrats!!!
- June 1, 2008 at 4:02 PM


text: Bleh. Not sure if I'm coming down with something or what, but this is the 2nd day in a row I've felt like crap. Achy all over, really tired, mildly depressed. Not majorly down, but lethargic, no motivation, no focus. There's plenty of stuff I know I should be doing, but I can't muster up the drive to do any of it. *Everything* requires too much conscious effort.


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"Pray always!" The Good Book tells us, and Jesus exhorted us to make sure that our prayer was patient, persistent, and persevering.
Eucharistic Adoration accomplishes this. It tells the world that "we can't give what we don't have" and that if we do not constantly turn to God in prayer for His grace and mercy, we're finished. The best thing people of faith can do is pray… I can't think of a better place to do that than before Our Lord in the Blessed Sacrament. (Timothy Cardinal Dolan, Archbishop of New York)
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