text:
L'opéra d'État de Vienne (en ) est une compagnie publique d'opéra et de ballet située à Vienne. C'est l'un des plus prestigieux opéras du monde, et l'une des toutes premières institutions culturelles de la capitale autrichienne. C'est aussi le nom du bâtiment inauguré en 1869 qui abrite cette compagnie

Histoire

Origines
La capitale de l'empire des Habsbourg, au croisement des mondes germanique, italien et slave, a très tôt été un centre européen de la musique : le premier opéra de l'histoire de la ville est donné en 1625 à la Hofburg ; l'Autriche développe au  une très riche tradition musicale. La Cour est le lieu de productions de spectacles dramatiques, lyriques et chorégraphiques de prestige, à la Hofburg même ou bien au Theater am Kärntnertor, une salle aux dimensions réduites qui s'étend bientôt aux bâtiments adjacents.

L'Opéra, en tant qu'institution, naît en 1810, avec la séparation des répertoires entre le Hofburgtheater, chargé du théâtre, et le k. k. Hof-Operntheater (Théâtre d'opéra de la cour impériale et royale), chargé de l'opéra et de la danse.

Installation sur le Ring

L'Opéra fut le premier des grands projets du Ring, et fut construit de 1861 à 1869. Les architectes, August Sicard von Sicardsburg et Eduard van der Nüll, le conçurent dans un style néorenaissance qui subit de violentes critiques. Il est vrai que la proximité du Heinrichshof, un hôtel, nuisait à son caractère monumental, et que le niveau de la rue devant lui avait été relevé d'un mètre après la construction ; il fut qualifié de . Van der Nüll se suicida, et Sicardsburg mourut d'un infarctus dix semaines plus tard, ce qui empêcha les deux architectes de voir leur ouvrage achevé.

Voir aussi la section Bâtiment.

Le nouvel opéra fut inauguré le  par une représentation du Don Giovanni de Wolfgang Amadeus Mozart (en allemand), peu de temps après la perte des provinces autrichiennes d'Italie du Nord.

Après une saison dans les deux salles, le Theater am Kärntnertor . L'institution sera appelée jusqu'en 1920 Kaiserlich-königliche Hofoper (Opéra de la cour impériale et royale, en abrégé k. k. Hofoper).

L'opéra des Habsbourg

La décennie de Mahler

Gustav Mahler favorisa l'émergence d'une nouvelle génération de chanteurs, avec notamment Anna Bahr-Mildenburg, Selma Kurz, Marie Gutheil-Schoderou Leo Slezak.

Il recruta également un nouveau décorateur, Alfred Roller, qui tourna le dos au style naturaliste et historiciste alors en vigueur pour créer des décors marqués par l'art nouveau.

Mahler favorisa la concentration sur l'œuvre en faisant baisser les lumières de la salle pendant la représentation, une innovation qui fut violemment critiquée mais maintenue par ses successeurs.

Des années 1910 aux années 1930
En 1920, l'Opéra prend son nom actuel, l'Autriche étant devenue une république.

La période nationale-socialiste
Le , un incendie ravage le bâtiment lors d'un bombardement américain ; la salle et la scène sont détruites, mais la façade, murée par précaution, reste intacte.

Les représentations reprirent dès le  mai au Theater an der Wien, alors inoccupé depuis longtemps.

L'après-guerre
Les formations de l'Opéra sont temporairement accueillies par le Theater an der Wien et le Volksoper.

En 1945, le chef d'orchestre Josef Krips fonda le Wiener Mozart-Ensemble, qui sous sa direction gagna une réputation mondiale en matière vocale et dramatique. Il comptait notamment dans ses rangs Erich Kunz, Elisabeth Schwarzkopf, et Wilma Lipp. S'éloignant de la tradition d'interprétation mozartienne avec grand orchestre, héritée du romantisme, Krips aida l'ensemble à développer un son léger et transparent proche de la musique de chambre et que l'on tiendrait plus tard pour typiquement viennois. Krips, qui commença là sa carrière mondiale, fut jusqu'à sa mort l'un des grands artistes de la compagnie viennoise.

Dès 1947, l'ensemble donne Don Giovanni à la Royal Opera House de Londres ; le rôle d'Ottavio était tenu par Richard Tauber, qui avait fui le nazisme et dut, trois mois avant sa mort, pour accomplir son rêve de chanter une dernière fois Mozart avec la troupe de Vienne, se produire avec un seul poumon.

Mais si l'Opéra avait reconstruit sa troupe, il était toujours en résidence dans une salle temporaire. La question s'est posée de reconstruire l'Opéra à l'identique ou bien de le raser et de construire une nouvelle salle, sur le Ring ou ailleurs. En 1946, le Chancelier Leopold Figl, notamment conseillé par le ministre de la Reconstruction Udo Illig, décide de l'ouverture d'un concours d'architectes. Le concours est remporté par Erich Boltenstern, avec un projet de reconstruction de la salle avec cependant une modernisation de la décoration dans le style de l'époque. Parmi les projets concurrents, Wilhelm Holzbauer avait notamment proposé un réarrangement de l'espace avec avancement de la scène et galerie supérieure faisant le tour de la salle, avec places aveugles.

Voir aussi la section Bâtiment.

La reconstruction, en une période de difficulté économique, est en partie financée par des dons privés, ainsi que par une collaboration avec l'Union soviétique qui fournit des matériaux.

Le , quelques jours après la proclamation de la neutralité autrichienne, l'Opéra rouvre avec une représentation de Fidelio de Beethoven dirigée par Karl Böhm, devant un parterre de personnalités autrichiennes et étrangères où comptait notamment le secrétaire d'État américain John Foster Dulles. L'ÖRF réalisa à cette occasion sa première retransmission en direct, alors qu'il n'y avait que huit cents postes de télévision dans le pays.

L'ère Karajan
Karajan inaugure ce que Christa Ludwig appellera « un nouveau style » de direction. Il amène avec lui un certain nombre de modifications, anecdotiques ou non : c'est lui qui décide de généraliser les productions d'opéras dans la langue d'origine, lui également qui inaugure des coproductions avec d'autres maisons, comme la Scala de Milan. De nombreux conflits sociaux éclatent sous son mandat ; certains sont dirigés contre lui, et d'autres le voient se ranger du côté des grévistes…

Le mandat de Karajan – qui s'achève en 1964 par une codirection ratée avec Egon Hilbert, qu'il détestait – reste dans l'histoire de l'opéra de Vienne comme le plus glorieux depuis celui de Gustav Mahler. Mais Karajan en paiera le prix ; estimant désormais impossible de maintenir une exigence de qualité tous les jours de l'année dans une maison d'opéra traditionnelle, il fondera, trois ans après son départ, le festival de Pâques de Salzbourg.

Des années 1960 aux années 1990

Aujourd'hui
En , le chef japonais Seiji Ozawa devient directeur musical de l'Opéra. Il avait dirigé le  de la même année le Concert du nouvel an de l'Orchestre philharmonique. À l'occasion des  de l'opéra, un écran haute définition de  est installé en face du bâtiment pour permettre la retransmission de l'intérieur vers l'extérieur. C'est le premier opéra du monde à proposer une innovation d'un tel genre. L'écran permettra, pour la saison 2009, la diffusion de plus d'une soixantaine d'œuvres.
À l'issue de la saison 2010-2011, première saison dirigée par le Français Dominique Meyer, le théâtre a réalisé

Statut

L'Opéra est une SARL (société à responsabilité limitée), appelée la « Wiener Staatsoper GmbH », filiale du Théâtre fédéral autrichien (le «  »).

Directeurs

La succession des directeurs est une longue liste de cabales et de claquages de portes :
 1867–1870 : Franz von Dingelstedt
 1870–1875 : Johann von Herbeck
 1875–1880 : Franz von Jauner
 1881–1897 : Wilhelm Jahn
 1897–1907 : Gustav Mahler
 1908–1911 : Felix Weingartner
 1911–1918 : Hans Gregor
 1919–1924 : Richard Strauss et Franz Schalk
 1924–1929 : Franz Schalk seul
 1929–1934 : Clemens Krauss
 1935–1936 : Felix Weingartner
 1936–1940 : Erwin Kerber
 1940–1941 : Heinrich Karl Strohm
 1941–1942 : Lothar Müthel
 1943–1945 : Karl Böhm
 1945–1954 : Franz Salmhofer
 1954–1956 : Karl Böhm
 1957–1964 : Herbert von Karajan
 1964–1968 : Egon Hilbert
 1968–1972 : Heinrich Reif-Gintl
 1972–1976 : Rudolf Gamsjäger
 1976–1982 : Egon Seefehlner
 1982–1984 : Lorin Maazel
 1984–1986 : Egon Seefehlner
 1986–1991 : Claus Helmut Drese, avec Claudio Abbado comme directeur musical
 1991–1992 : Eberhard Waechter
 1992–2010 : Ioan Holender, avec Seiji Ozawa comme directeur musical à partir de 2002
 Depuis 2010 : Dominique Meyer, avec Franz Welser-Möst comme directeur musical (2010-2014)

Formations

Orchestre
L'Orchestre de l'Opéra () est très lié à l'Orchestre philharmonique de Vienne () : les  forment une association indépendante, mais ne recrutent que parmi les musiciens membres de l'Orchestre de l'Opéra depuis plus de trois ans. Ceci garantit à l'association, qui ne reçoit pas de subvention, son indépendance financière, car ses musiciens reçoivent déjà un salaire à l'Opéra.

C'est l'Orchestre de l'Opéra, et non l'Orchestre philharmonique, qui se produit à l'Opéra ; en revanche, c'est l'Orchestre philharmonique qui se produit au Festival de Salzbourg et réalise des enregistrements en studio.

Il compte parmi ses anciens premiers violons Arnold Rosé, Wolfgang Schneiderhan et Willi Boskovsky.

Chœur
Le Chœur de l'Opéra (Wiener Staatsopernchor) participe à des enregistrements et à d'autres manifestations musicales, en premier lieu le Festival de Salzbourg, sous le nom de Konzertvereinigung Wiener Staatsopernchor (Association de concert du Chœur de l'Opéra d'État de Vienne).

Bâtiment

L'Opéra est l'un des bâtiments les plus connus de Vienne, où il est appelé la erste Haus am Ring (« première maison du Ring »). Le bâtiment d'origine de August Sicard von Sicardsburg et Eduard van der Nüll, construit dans le style néoromantique et sévèrement critiqué lors de sa construction, fut inauguré le . Il s'appuie sur les styles du passé, notamment de la Renaissance pour les arcs de la façade.

Pendant la Seconde Guerre mondiale, le , la scène fut détruite par un bombardement allié et le bâtiment ravagé par un incendie, à l'exception du foyer, du grand escalier, du vestibule et du salon de thé. La quasi-totalité des décors, des accessoires de plus de cent vingt opéras et de plus de cent cinquante mille costumes fut détruite.

L'Opéra fut reconstruit après la guerre par Erich Boltenstern. Sur le conseil d'Arturo Toscanini, il employa beaucoup de bois afin d'améliorer l'acoustique. Le nombre de places au parterre fut réduit et la cloison autour du quatrième balcon supprimée, la capacité de la nouvelle salle étant de deux mille deux cents places. La façade, l'entrée et le foyer décoré par Moritz von Schwind, épargnés par l'incendie, furent restaurés et gardèrent leur style premier.

Fonctionnement

L'Opéra fonctionne selon le système dit du répertoire : il entretient des ensembles permanents, à traitement fixe, connaissant un certain nombre de productions déjà rodées, qu'il fait se succéder à l'affiche. Ceci lui permet de programmer lors de sa saison une cinquantaine d'opéras et une vingtaine de ballets, tous les soirs du  septembre au , sauf le  et le Vendredi saint : plus d'une dizaine de spectacles peuvent se succéder à l'affiche en deux semaines. Cette méthode, bien qu'elle offre une grande variété au public et lui permette de se former, est cependant susceptible de nuire à la qualité des spectacles, peu préparés et donnés dans une certaine routine.

Le Bal de l'Opéra
Le Bal de l'Opéra (Opernball), le plus prestigieux de la saison des bals, a lieu chaque hiver le jeudi précédant le mercredi des cendres. Il se tient dans la salle des spectacles, le parterre étant rehaussé jusqu'au niveau de la scène pour servir de piste de danse. L'événement rassemble des personnalités de la haute société, du monde des affaires et de la politique.

Depuis quelques années, des mouvements d'extrême-gauche organisent une Opernballdemo (« manif du Bal de l'Opéra ») le même soir sur le Ring, afin de protester contre les valeurs incarnées par certains visiteurs richissimes du bal ; elles ont parfois donné lieu à des violences. L'écrivain Josef Haslinger, dans son roman Opernball (1995), a pour sa part imaginé le massacre de l'élite du pays par un attentat commis par des néo-Nazis

Galerie de photographies

Voir aussi

Bibliographie
 Rudolf Klein, L'Opéra national de Vienne. Un guide à travers la maison et ses cent ans d'histoire, traduit de l'allemand par Catherine Foramitti, Verlag Elisabeth Lafite, Vienne, 1967
  Marcel Prawy, Die Wiener Oper. Geschichte und Geschichten, nouvelle édition revue et augmentée, Verlag Fritz Molden, Vienne, 1969
 Andrea Seebohm (direction), L'Opéra de Vienne (1986), traduit de l'allemand par Odile Demange, Seuil, Paris, 1987
 « Vienne, à », Le Monde de la musique, hors-série « Karajan, la légende du chef », 1999

Articles connexes
 Vienne (Autriche)
 :Catégorie:Culture autrichienne
 Orchestre philharmonique de Vienne

Liens externes

   Programme de l'Opéra d'État de Vienne
  Mahler et l'Opéra de Vienne, sur le site de Martijn Hooning

Notes et références

Notes

Références

Salle d'opéra à Vienne (Autriche)
Musique en Autriche
Architecture civile du XIXe siècle
Patrimoine du XIXe siècle
Culture à Vienne (Autriche)
Édifice construit en 1869


text:
[![Build Status](https://travis-ci.org/77Vincent/toggling.svg?branch=master)](https://travis-ci.org/77Vincent/toggling)
[![License: MIT](https://img.shields.io/badge/License-MIT-yellow.svg)](https://opensource.org/licenses/MIT)

Element's state toggling in browser

## Demo
Demo page

## Installation
##### NPM

```sh
npm install --save toggling
```

##### CDN

```html
<script src="https://raw.githubusercontent.com/77Vincent/toggling/master/toggling.min.js"></script>
```

## Instantiate

```js
import Toggling from 'toggling';
//or
var Toggling = require('toggling');

new Toggling({
    trigger: 'trigger-selector',    // required
    target: 'target-selector',      // required
    handler: function(tar, tri) {   // required
        this.toggle(tar);
    },
    excluded: 'element-selector',   // optional
    event: 'click'                  // optional
    useCapture: false               // optional
});
```

## API

### Configurations
##### trigger: String
* Multiple elements will be returned if found, it's fine if no element is found.

##### target: String
* Only one single element will be returned if found, it's fine if no element is found.
* Will be first looked for elements that either are adjacent or descendant to the trigger element.
* Target could also be declared in trigger element in markup:

```html
<div data-toggling="this-is-target-selector">This is trigger element</div>
```

##### excluded: String
* Element that will be excluded from triggering the event
* Only accept one selector.
* Will be only looked for elements that either are adjacent or descendant to the trigger element.

##### handler: Function
* There are 2 default passed arrguments to be accessed: tar, tri

```js
handler: function(tar, tri) {
    // tar is the found target element
    // tri is the current trigger element
}
```

##### event: String
* Event to trigger the handler
* Default: click

##### useCapture: Boolean
* If capture or bubbling mode will be used.
* Default: false

### Instance functions
##### disable()
* Remove all event listener from triggering elements.

```js
var toggling = new Toggling({
    // configurations...
});

toggling.disable();
```

* Or could be called inside handler:

```js
new Toggling({
    trigger: ...,
    target: ...,
    handler: function() {
        this.disable();
    }
});

// This instance will be disabled after being triggered once.
```

##### enable()
* Bind event listener to triggers again.

```js
toggling.disable();
// Let's say you disabled an instance first

toggling.enable();
// Then you can enable it again.
```

##### show(element)
* Display the passed element.

##### hide(element)
* Hide the passed element.

##### toggle(element)
* Toggle the passed element.

##### addClass(element, className)
* Add className to element.

##### removeClass(element, className)
* Remove className from element.

##### toggleClass(element, className)
* Toggle className from element.

##### closest(element, className)
* Find the closest ancestor element to the passed element, according to the passed className.

## Compatibility
IE 9+

## Motivation
As we have a lot of pop-up features, the core concept behind is state changing. The goal of creating this module is to giving a simple, robust and versatile solution for this kind of feature. So that developers don't need to write similar codes repeatly.

The state changing model could be abstracted as "Trigger-Event-Handler-Target".

Besides, sometimes we don't want a certain child element to trigger the event, for instance we want to close an pop-up box by clicking anywhere on the screen but not a certain area, in toggling, then we can bind event to 'html' element, which represents the whole document, then exclude the certain element from event triggering.

What's more, each toggling instance provides you some very basic DOM-manipulation methods like show or hide element, add or remove class, which work without any javascript frameworks. But if you want to achieve more fancy effect in your handler, maybe jQuery, Prototype.js or some other javascript frameworks would be the best partner.

redpajama_set_name: RedPajamaGithub


A mold and mildew assessment will identify any problems around your residential property which will sustain mold and mildew growth as well as pinpointing problem areas for instance water damages, smells as well as evident indications of fungal growth and so on. The mold assessor ought to take the inspection outside as well as examine the outside of the dwelling once more for obvious signs of water damages, leaks as well as their sources such as busted pipes, damaged seamless gutters, fractured brick job and so forth.
As a rule of thumb, these examples can take 3-4 days to refine at the lab however I would certainly expect that your mold assessor make get in touch with as well as offer a verbal record as quickly as the outcomes become available adhered to by a written record within a number of days. An added point to be aware of, when it concerns the report following a mold examination a great mold and mildew examiner will certainly not merely send you a duplicate of the lab record. He/She has to able to confidently translate the results and supply them along with suggestions to resolve the concern !
NB. Need to you reserve a mold examination it is very important that you keep doors and windows shut as long as possible for the 12 to 24 hrs before the assessors check out. This will certainly permit that the mold inspection results are exact. It will enable the mold and mildew inspector to detect smells as well as air examples will not contain excess outside air.


text:
Lessons from Amazon's Italian hub strike: industrial action that does not factor in both work AND data is doomed to be ineffective
28 November 2017 adminBlogamazon, amazon mechanical turk, digital labor, platform, strike, unions
On Nov 24, 2017, the three main Italian unions (CGIL, CISL, UIL) have called for a strike over the failure to negotiate Black Friday bonuses for the 1,600 permanent workers at the distribution hub near the Northern town of Piacenza. Unions say 50% of the workers partake in the strike. Amazon says it was more like 10%.
Bottom line: the strike did not stop Black Friday in Italy. Someone was working. Yet, according to several sources, it was not not permanent workers, but the 2,000 temps that Amazon recruited until Xmas who saved the day. They were not hired to replace striking workers. Even in Italy, this would be illegal. They were hired to face Nov./Dec. surge in retail sales. And of course they did not stop working on Black Friday 2017. That said, Amazon is known internationally for its brutal workplace discipline, its anti-labor stance, and has been accused of hiring temps, contingent workers and even workampers to edge out unionized labor force.
In Italy, one can recruit a lot of those. Unemployment is at 11.1% and there's a millions-strong industrial reserve army of faux-freelance, part-timers, "coordinated collaborators", "project-contractors", "leased staff" and many other forms of non-standard employees. Especially since the infamous Jobs Act heralded by the government of former PM Matteo Renzi, among young workers temp jobs accounts for 50% of employment and they are up 7% since Sept 2017.
But Italian retail workers and their strike tell only part of the story. Amazon isn't about e-commerce: it's about big data. Interestingly, Matteo Renzi's government has been very helpful in facilitating the strategy of "data entryism" of the Seattle giant, going as far as to hire Amazon's former vice-president and now-biggest employee shareholder of the platform as "Commissioner for Digital Italy". He's doing this for free, and you know what they say when you're not paying for something…
Which brings us to the main point. Amazon strategy is predicated on data and work. Even better: it is predicated on data-as-work, because it extracts value from the data stored in its humongous cloud and hosting services, and because it uses people-as-a-service (according to Jeff Bezos's early characterization of Amazon Mechanical Turk) to train, enrich, refine data.
Btw, do you wanna know what the new Italian Digital Commissioner considers as a success story for digital transformation? The controversial Indian biometric ID system… And do you know where 36% of Amazon Mechanical Turkers live? India… (Here's the interview [in Italian] where the Digital Commissioner talks about Indian ID system while at the same time declaring that "he misses Amazon so much").
Take-away message: Amazon corporate takeover of Italy is as much a matter of labor policy as it is of data politics. As long as the unions continue to focus on the former while neglecting the latter, their action is doomed to be ineffective. Case in point: after dominating Black Friday sales, Amazon's shares are up 2% and Jeff Bezos is still world's wealthiest man. So Amazon Italia just gave a giant middle-finger to workers by cancelling the meeting with unions and rescheduling it for after Xmas…
Colonialisme numérique ? (dans Usbek&Rica, oct-nov. 2017)
"I lavoratori sono il cuore dell'algoritmo" : intervista ne Il Manifesto (Italia, 9 dicembre 2017)

redpajama_set_name: RedPajamaCommonCrawl


How Will A Bear Market Affect Your Plan?
In this week's edition of the Ballast Atheneum, I am going to explore a subject no one wants to see but we all must prepare for; Will my portfolio survive a bear market? Fortunately, we can use some of our technology tools, specifically, Monte Carlo analysis to determine how prepared a client is for a bear market.
It is no wonder we witnessed a spike in mutual fund sales during this period. What proved to be the investment opportunity of a lifetime drove many investors to the sidelines. Sadly, many people let fear dictate their decision making and sold near market lows. Conversely, those who were able to stick to a long-term allocation plan and had confidence in that plan, would have recognized that period as a reality of being a long term investor.
As investors, particularly those accumulating investment assets for a certain goal (i.e. retirement), we must understand that down market periods are inevitable. We have discussed in recent commentaries that we believe market sell-offs and increased volatility are part of healthy markets over the long term. However, we do not believe the best approach to dealing with this inevitability is to just sit on your hands. We have consistently encouraged our clients to be proactive in ANTICIPATION of bad markets, not in REACTION to them. Furthermore, we do not believe this action is one based in short-term thinking, it is critical to think of such positioning in the context of your long-term plan. We spend a great deal of time with our clients figuring out what allocation fits their long-term goals, knowing that they will experience markets, both good and bad.
Is your portfolio positioned to deal with inevitable market pullbacks? The good news is we have tools to make more educated decisions when trying to answer this question.
As stated above, each achieved the exact same average return over a relatively short term period of five years and the final results are drastically different.
-Bill ends the five years period with over $314,000 less than Julie. Notice how his first two years were negative.
-Even with only one poor year to start her plan, Mary ended her plan with over $100,000 less than Julie.
-The most desirable result came from Julie with steady returns of 8% each year.
Although Julie's "sequence" is virtually unattainable, this example does quickly explain why a portfolio that achieves steady returns (misses big up years and avoids big down years) is most desirable for many investors. If we look at Bill, his portfolio could resemble someone who was an aggressive investor leading into the financial crisis. Bill experienced two poor years of market returns and even with three very positive years to follow, he still is far behind the other three portfolio examples.
When we sit down with investors who are planning to take portfolio distributions, we prefer to run long-term Monte Carlo analysis to display what impact a poor sequence of returns might have. Below is an example of a plan showing portfolio income when someone achieves the same return each year.
As I mention above, markets have rarely ever provided equal returns over a five year period but unfortunately this is how most people view their financial plan. In this next example, a period similar to the Financial Crisis is applied in the early years of the plan. Notice the difference in long-term preservation of assets when the household experienced negative returns in the first couple years of retirement.
You can see in the above graph that the portfolio survives both lifetimes but the ending values are VASTLY different. For the purposes of this example, I am using a portfolio that is 100% invested in stocks. In the next chart, when we shifted the allocation to one with other asset classes (including 25% in bonds) and displayed increasing 401k savings through retirement by 4%, we achieved a much more desirable result.
The above chart is a great example of the eventual "compromise" we make with our client plans. A client may have expectations of retiring at a certain age on a certain budget and our final plan recommendations often include changes to the budget, allocation, retirement date, or savings up until retirement. In each scenario we see, slight changes to any of the factors can have an impact on our final Monte Carlo projections.
We can test your current and expected future portfolio to determine if your plan can withstand a poor sequence of returns.
By shifting your allocation and increasing your savings up until retirement, we can increase your chances for success.
We prefer to make portfolio allocation changes well in advance of retirement: at least several years in most cases.


Such striving may seem admirable, but it is a way of endless frustration for the unobtainable, trying to please the 'unsatisfiable' in a craving world.
Help them instead to find the wonder and the marvel of life as God makes it.
Show them the joy of tasting home grown tomatoes, apples, pears, figs.
the crawl of a crab in an ocean pool, the delight of a dog at play.
that our kids love us and we eat well enough to enjoy simple pleasures, bush walking, gardening, bird watching, beach combing, rock climbing.
and guiding us through the loops and twirls.
Having a strong will is good. We want our child to have a strong will.
to choose eternal faith, a strong will to swim upstream, etc.
We need to guide their strong will for wisdom for their good.
Never discipline if angry - quickly pray for calmness.
so there is not a battle of wills constantly.
but the years after that are much sweeter.
The family can go out more, do more things, sleep anywhere, if they know they are safe in parents care.
for they know the parent's heart and feel safe in the training they have received.
They will come when called, they will stop when danger threatens.
ie Discipline is only so that they know survival skills.
- crossing roads, dangers, manners etc.
They must receive a just punishment for deliberate rebellion or they will carry the guilt of it for life.
A punishment releases them of the burden of carrying the disappointment in themselves.
(practical tip -only on the fat of the legs where it will not bruise - with eg a wooden spoon until they whimper,ie if they are still angry and thrashing with rebellion you have not conquered the rebellion.
When the child is crying softly -we must then hug them and assure them of forgiveness and love.
a natural instinct to protect the little one.
I didn't make rules for them to have to rebel against. They grew to know how life worked.
and they worked it out through discussion.
Thank you, Dr. Dobson. Your books and advice helped us all through our child rearing years. I read The Strong Willed Child. and applied the principles. They are all married with children of their own - a testimony to these truths.
and He will lead them through trials of life.
We CAN trust Him, He loves them more than we do.
We can but watch, pray, suggest or/and wait for, if they need help.
she did not attempt to molly coddle /protect/explain away/deny or shield him from life's realities.
Franklin became a great man.
Sometimes we can 'parent' for our own reputation.- that we may look good.
It never works, the child knows that they are not genuinely loved.
That is not loving the child, that is loving self.
just as God is always there/with us, to mould/direct us to wise living.
we are weak and need God, we cannot breathe without HIM.
that we will see them safe eternally.
loves them and has a home for them forever.
Parents have your children experienced tenderness and mercy in your home today?
Parents can only show loyalty to God when loyal themselves. A child trusts only what is true.
without breaking his spirit and damaging him in some way?
through Christ we are redeemed/set free from the power of our sinful heart.
It is not through our own effort/try/try but only through the Holy Spirit that we can "do" this.
He will give us wisdom and strength to choose HIS way/His heart, when we trust Him.
a parents prayers should continue until we draw our last breath - pray for their future spouse when they are young. Ask the Lord to prepare both your child and the mate He has for them.
What are the long-range implications of raising a strong-willed child?
What can we expect as the years go by?
Well, I can give you a few encouraging conclusions from our study.
The tendency of strong-willed children is to return to parental values when they reach adulthood.
Parents told us that 85 percent of their grown SWCs (twenty-four years of age and older) came back to what they had been taught--entirely or at least "somewhat." That is good news.
Only 15 percent were so headstrong that they rejected their family's core values in their mid-twenties.
In those exceptional cases, I'll wager that other problems and sources of pain were involved.
but the majority will turn around when they reach young adulthood and do what their parents most desired.
That should be reassuring. Furthermore, if we could have evaluated these individuals at thirty-five instead of twenty-four years of age, we would have seen that even fewer were still in rebellion against parental values.
Second, raising a strong-willed child (or a houseful of them) can be a lonely job for parents.
You can begin to feel like yours is the only family that has gone through these struggles. Don't believe it.
we found that 85 percent of families had at least one strong-willed child.
God's gift to us. Beautiful and innocent -Remember where they came from and treat them with respect and lots of LOVE..
" MAKE OR BREAK "
A high school student said that he was stupid. I asked, "What makes you think that? "
He said, "My dad says that I am." He believed what his father said.
He was NOT stupid, but his grades were pretty bad.
He had leadership potential, but he was "leading" in the wrong way.
He didn't try for good grades because he thought that he was "stupid".
I couldn't convince him that he was smart. It's hard to compete with a parent.
I wonder what I could have done.
PARENTS, YOU HAVE THE POWER TO " MAKE OR BREAK " YOUR CHILD.
He could very easily become the child that you tell him that he is!
Everyone is different, what you think is clever about you, will not be the same in your child.
God has made them so.
You do what you need to do, they will do what they need to do.
Correction is one thing, belittling them to embarrassment is another.
that just a 'look' or a shake of the head will adjust the child's choices.


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Three Films by Jia Zhangke (15.)
Directed by Jia Zhangke.
A three-disc, dual format Blu-ray/ DVD box set released by Arrow Academy including:
Mountains May Depart. (12A.) 2015. Starring Tao Zhao, Yi Zhang, Jing Dong Liang, Zijian Dong, Sylvia Chang, Sanming Han. 123 mins.
A Touch of Sin. (15.) 2013. Starring Wu Jiang, Vivien Li, Lanshan Luo. 127 mins.
24 City. (U.) 2008. Starring Joan Chen, Tao Zhao, Jianbin Chen, Liping Lu. 109 mins.
Around a decade ago some fairly outlandish claims were being made for the work of Jia Zhangke, some reviewers calling him one of the world's most important filmmakers. Well, I don't know about that, but he specialises in one of the world's most important and fascinating topics: the consequences of China's mass lurch into state capitalism. For international audiences he has been the Chinese filmmaker who has best illustrated the dramatic social and political upheavals in his country during this century. With his wry, detached style he has captured the ironies of a one-party state unleashing the forces of unchecked capitalism, the brutalities of industrialisation and the pains of a rural society being hurried along into becoming urban high rise one.
His filmmaking has mirrored that shift. He started out as an arthouse director/ documentary maker. I first encountered his work at a London Film Festival about a decade ago with the film Still Life, a quiet taciturn affair, more documentary than drama, about people being displaced by the Three Gorges Dam project. From there he has gradually moved away from the margins towards something more mainstream: films with stories and characters where occasionally things happen. Whereas his films used to be things that would appear at film festivals and get a brief run out down at the BFI Southbank, now they will appear in a more prominent position at film festivals and get a slightly longer run out down at the BFI Southbank and be shown at a few other arty cinemas in selected metropolitan areas across the land.
The landscape of his filmmaking is filled with half-built airports and bridges, gleaming train stations, communities scattered across the hillsides surrounding a lake or reservoir, workers dormitories and motorbikes moving through dust. There is the occasional neon-lit city street but mostly he concentrates on small (for China) communities and the harsh terrain. The landscape dominates the characters, though over the course of this collection his filmmaking becomes more focused on the performers.
His work is quiet and understated. He's not much of a dramatist and I suspect he doesn't much like working with actors (well, why would you?) but an ability to frame truly telling images allows him to really gets to the heart of life in China. It's a society in constant and unruly flux, families and relationships being pulled apart as people migrate across vast distances to find menial, low paid work in factories and mines. It's as if Mao's Long Marsh had never really but simply dissipated into a nationwide scramble, a billion people zigzagging across the country trying to find a place for themselves.
Arrow's box set gathers together his last three features films, a decade of work. (Outside of these he makes shorts and documentaries.)
24 CITY.
A TOUCH OF SIN.
MOUNTAINS MAY DEPART.
Extras.
The Extras are very much a Tony Rayns joint, the film critic who has specialised in East Asian cinema. He provides quarter hour introductions for each film (probably best seen afterwards) and conducts two of the three lengthy interviews with the director that are on each disc.
There are also a couple of Jia Zhangke's short films, some Making Ofs, a booklet, a visual essay placing Jia Zhangke work in the context of Chinese cinema.
Plus, there's a Limited Edition exclusive: a full length (105 mins) documentary on him by the Brazilian director Walter Sallas (Motorcycle Diaries, On The Road.) All of these reveal Jia Zhangke to be a short, owlish fellow, resembling Andre Previn.


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The Model Institutions for Excellence (MIE) Program Report
The Model Institutions for Excellence (MIE) program, initiated in 1994, is a joint venture between the National Science Foundation (NSF) and the National Aeronautics and Space Administration (NASA). The program was designed to increase the number of underrepresented minorities in science, technology, engineering and mathematics (STEM) through funding to a select group of minority-serving institutions (MSIs).
The MIE program aims to increase the representation of minorities in STEM by:
Targeting a small number of MSIs poised to make a substantial contribution to increasing the number of minorities who earn STEM baccalaureate degrees and then enroll in STEM graduate programs or enter STEM careers;
Improving STEM education and undergraduate research at the selected MSIs; and
Enabling successful projects to serve as models for the recruitment, education and production of quality-trained STEM baccalaureate degree recipients.
In 1994, 69 MSIs were invited to submit MIE planning proposals. Fifty-seven MSIs submitted proposals and 20 were funded to develop implementation proposals. Six projects (one of which is a consortium of three colleges) received long-term funding for infrastructure development in STEM education and individual support to recruit and retain minority STEM students. The six MIE projects include:
Universidad Metropolitana in Puerto Rico;
Xavier University of Louisiana;
University of Texas at El Paso;
the Oyate Consortium (composed of Oglala Lakota College, Sitting Bull College and Sisseton-Wahpeton College located in South and North Dakota);
Spelman College in Georgia; and
Bowie State University in Maryland.
Xavier University, Universidad Metropolitana, University of Texas at El Paso and the Oyate Consortium are funded by the National Science Foundation. Spelman College and Bowie State University are funded by NASA.
Highlights from the Study
With one exception, from 1997-98 to 2003-04, STEM enrollment tended to increase faster than overall institutional enrollment at each MIE institution. (At Universidad Metropolitana, the enrollment more than doubled during the period, from 3,294 to 7,499 students (a 128-percentage point increase) and although STEM enrollment also more than doubled, it did not quite keep pace with the total.)
With one exception, from 1997-98 to 2002-03, the number of undergraduate STEM degrees conferred and the proportion of all degrees awarded that were in STEM fields increased considerably in all MIE institutions.
STEM degrees awarded tended to increase faster in the MIEs than they did overall in the Historically Black Colleges and Universities (HBCUs), Hispanic-serving Institutions (HSIs), and in the group of non-funded MIE applicants.
There appears to be one MIE model with seven essential components: recruitment and transition initiatives, student support, undergraduate research, faculty development, curriculum development, physical infrastructure development, and STEM graduate school and employment initiatives. Although each project looked somewhat different, student support (including social, financial, and academic assistance) received significant emphasis across all projects. Infrastructure enhancements included improvement or development of classrooms, laboratories, and specific areas in which students could study and work; purchases of state-of-the-art computing and laboratory equipment; hiring of over 100 new STEM faculty; curriculum enhancements at every project; and new STEM degree programs established at many. Undergraduate research opportunities were available both on- and off-campus, anchoring the students' motivation and persistence in STEM.
The study suggests that the MIE model is readily transportable, but that it must be aligned to the context and culture of the institution.
Creating and Maintaining Excellence: The Model Institutions for Excellence Program Report
Increasing Underrepresented Minorities in STEM Education and Professions
In 2004, NSF contracted with the American Institutes for Research (AIR) to conduct a short-term study to assess program impact and collect evidence of project success in infrastructure enhancement, and student recruitment, retention, graduation and advancement in STEM careers with an emphasis on achieving and sustaining diversity in the STEM ...


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Ганс Цах (;  30 березня 1949, Бад-Тельц, Баварія, ФРН) — колишній німецький хокеїст, нападник. Згодом хокейний тренер. Член Залу хокейної слави Німеччини.

Ігрова кар'єра
Свою кар'єру почав у команді «Бад Тельц», відігравши три сезони перейшов до СК «Ріссерзеє». У складі СК «Ріссерзеє» Ганс відіграв чотири сезони, ще сумарно десять сезонів відіграв за СК Берлін, «Ландсгут» та СБ Розенгайм.

У складі національної збірної відіграв на чотирьох чемпіонатах світу: 1969, 1976, 1977 та 1978, а також на Зимових Олімпійських іграх 1980.

Тренерська кар'єра
З 1985 по 1987 тренував молодіжний склад СБ «Розенгайм». У 1987 році закінчив тренерські курси та очолив у 1990 Дюссельдорф ЕГ з яким чотири рази поспіль здобував звання чемпіонів Німеччини.

Також очолював ЦСК Лайонс, «Кассель Хаскіс», «Ганновер Скорпіонс» (золото 2010) та «Адлер Мангейм».

Залучався, як асистент головного тренера національної збірної на Кубок світу та Олімпійські ігри 2002.

Статистика

Клубна

Збірна

Посилання
Профіль на hockeydb
Профіль на eliteprospects
Профіль на eurohockey
Профіль на sports-reference

Німецькі хокеїсти
Хокеїсти «Бад Тельц»
Хокеїсти СК «Ріссерзе»
Хокеїсти СК «Берлін»
Хокеїсти «Ландсгута»
Хокеїсти «Розенгайма»
Німецькі хокейні тренери
Хокеїсти на зимових Олімпійських іграх 1980

redpajama_set_name: RedPajamaWikipedia


Are you a member of the Institute of Advanced Motorists?
Do you have a criminal record? Do not include parking offences.
If unsure, just put an estimate.
Is your vehicle SVA passed?
Is your vehicle left hand drive?


If you feel that you have been exposed to asbestos – what do you do? Any recent exposure to asbestos within the last ten years should not cause immediate symptoms or illness.
If you are in paid work and you feel that you have been exposed to asbestos then you need to report this to your employer. Your employer should then note the date, place and circumstances of your exposure in your personal file.
If you belong to a trade union, you should contact your union for advice and they will probably ask you for information which will also be recorded. This is for your benefit.
It is also be advisable to inform your GP that you have had an asbestos exposure and ask that it be recorded in your medical records – this is in case you should go on to develop an asbestos related disease.
Please get in touch with us here at AASC – Tel: 01495 272 479 or email enquiries@a-a-s-c.org.uk if you feel that you have been exposed to asbestos. We can provide you with help and support .
Some of the occupations which may have entailed exposure to asbestos.
Asbestos fibres are so fine that they can cling to clothing. It is good practice to change out of work clothing before arriving home, but in the past it was very common for workers to bring home their overalls after work each night. These would then be picked up by the wife usually and washed. If this describes you and what happened in your life or someone you know, bear in mind that if a person worked with asbestos it would be very likely that asbestos fibres would have contaminated clothing. Whilst the overalls were being handled through to the washing process the particles of asbestos dust could be inhaled.


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Busting Copenhagen's phantom 0
From Investors.com (Investor's Business Daily):
With less than two months to go before the big Copenhagen Conference on global warming, two major nations have said "no thanks" to the no-growth agenda. For that reason alone, so should we.
Following a deal signed late Thursday between China and India, anything we might agree to do in Copenhagen is likely moot anyway. The two mega-nations — which together account for nearly a third of the world's population — said they won't go along with a new climate treaty being drafted in Copenhagen to replace the Kyoto Protocol that expires in 2012.
They're basically saying no to anything that forces them to impose mandatory limits on their output of greenhouse gas emissions. Other developing nations, including Mexico, Brazil and South Africa, will likely reject any proposals as well.
The deal was already in trouble. Three weeks ago, the Group of 77 developing nations met in Thailand to discuss what they wanted to do about global warming. Their answer: nothing. …
They see clearly what the rest of us seem to miss — that, for all its bad science, the Copenhagen Conference is about the world's Lilliputians tying down its Gullivers, not about global warming at all.
So, thanks to China and India, Copenhagen is dead — just as Kyoto was when it was signed in 1992, though no one knew it at the time. Without them, no global treaty on climate change will be workable.
The two nations are not only the world's most populous (with, together, more than 2 billion people), they are also the fastest-growing major countries. China is now the world's No. 1 emitter of greenhouse gases, and India is catching up fast.
Even with their participation, Copenhagen should have been a non-starter for the U.S. Indeed, the main reason for the greenhouse gas deal, all but admitted to by its major participants, is to cripple the U.S. economy — the most successful economy in the world.
True enough, as green critics keep saying, we produce nearly 20% of the world's CO2 and other greenhouse gases with just 5% of the world's population. But our GDP of roughly $14 trillion is nearly 25% of the world's total — in line with our gas output.
We provide jobs and consumption not just for Americans, but for tens of millions of people overseas whose livelihoods depend on satisfying the massive American market.
In case you're still worried about warming, stop. Since 1998, the data show global temperatures have fallen. The Intergovernmental Panel on Climate Change says this can't be happening. None of the IPCC's models shows a possibility of rising CO2 output and declining temperature.
But even Paul Hudson, the pro-warming-theory BBC climate correspondent, recently had to admit: "For the last 11 years, we have not observed any increase in global temperatures. And our climate models did not forecast it, even though man-made carbon dioxide, the gas thought to be responsible for warming our planet, has continued to rise."
Yet, the IPCC estimates that "remediation" of the warming trend will cost about 1.7% of world GDP. In the U.S., that's about $240 billion a year. For the entire world, it's about $1 trillion a year — or $71 trillion over the next 70 years or so.
Proposals to slash CO2 won't work anyway. Department of Energy estimates indicate that 97% of all CO2 emissions would continue even if humans didn't exist.
Even so, climatologist Chip Knappenberger estimates that laws like the recent Waxman-Markey bill would, if fully enacted, reduce future warming by just 0.2 degrees Celsius by 2100 — not enough even to measure accurately.
Can the world really afford to give up $71 trillion in the coming decades to solve a phantom problem?
Given the shoddiness of the science behind warming claims and the refusal of the biggest CO2 emitters to play along with the climate change sham, it would be economically ruinous for the U.S. to do anything other than wish the rest of the world a nice day, and go about our business.
Posted under China, Climate, Economics, Environmentalism, India, News, United States by Jillian Becker on Saturday, October 24, 2009
Tagged with China, Chip Knappenberger, climate change, CO2 emissions, Copenhagne Conference, Global warming, India, IPCC, Waxman-Markey


President Obama signed an executive order raising the minimum wage for many federal contract workers starting in 2015.
President Obama mandated the increase, from the country's federal minimum wage of $7.25, by signing an executive order on Wednesday.
The new $10.10 hourly wage will be indexed to inflation after the first year, and it will apply to contracts and subcontracts that provide the federal government with concessions, services and construction. It will not, however, cover workers employed by businesses that provide goods, such as supplies, to Uncle Sam.
The move comes after a push from liberals, who worry that lawmakers will not move quickly, if at all, on legislation to gradually raise the minimum wage for everyone to $10.10 an hour.
Obama's order will likely affect fewer than half a million workers.
But the hope is it will spur Congress to approve the same increase for all workers.
Raising the minimum wage, the White House and other advocates argue, would have many positive effects, including increased productivity and lower turnover. Just as importantly, they say, it can lift a family of four supported by a full-time minimum wage worker out of poverty.
"Not only is it good for the economy, it's the right thing to do. You work full time? You shouldn't be living in poverty," Obama said.
The president's order will also apply to disabled workers employed by federal contractors. Currently they may be paid less than the federal minimum wage under certain certificate programs.
Tipped workers employed by businesses with federal contracts would also benefit. Currently workers who receive tips are paid a minimum required base of $2.13 an hour.
Obama's order will raise that base to $4.90 and increase it by 95 cents a year until it reaches 70% of the regular minimum wage. If a worker's tips on top of that don't add up to $10.10 an hour, the federal contractor will be required to make up the difference.
Among the types of workers who would benefit from the president's executive order are concession workers in national parks, nursing assistants caring for veterans, people who serve food to U.S. troops and those who maintain the grounds on military bases, the White House said.
Critics say a higher minimum wage will push employers to hire fewer workers or reduce their hours.


If you've been wondering about the SAT, you need wonder no more. Here we feature the best ways to SAT Prep, online SAT prep, SAT prep classes, SAT prep books, and more. So, let's get started!
Many colleges use the SAT in combination with students' GPAs to make admission and scholarship decisions.
The higher your SAT score, the more likely you are to get into your desired college and/or get a good scholarship. According to Fred Zhang, a top SAT tutor, your SAT score impacts 30 to 50% of your dream college's admission's decision.
From experience, I can tell you this is true. In high school, I was in the bottom half of my class. My grades weren't fantastic. However, I scored a 2160 on my SAT in 2013, which today would equate to a 1500. I was in the top 1% of all test takers. When I applied to colleges, I was admitted into some selective technical schools. Also, I was offered scholarships equaling half my tuition every year. Not bad, right?
For some lists of guaranteed scholarships based on SAT scores, check here, here, here or search "SAT Scholarships".
Also, the United States military offers scholarships with their ROTC programs. These can depend partially on SAT scores. ROTC is a good way to pay for college if you're willing to serve in the military after college.
So, to put it plainly, it is in YOUR best interest to score as highly as you can on the SAT. The rest of this guide will teach you the best ways to SAT prep.
To score highly on the SAT test, you're going to need internal motivation. This means that you want to achieve something just for the purpose of achieving it. You value the long-term benefit, not immediate gratification. You're motivated by the big picture reward.
So how do you get motivated?
Picture in your head what achieving that desired SAT score would look like.
You log into your account online, and waiting for you is that juicy, high score. Bam! You know that your hours of studying have paid off and that you've got a good grasp of the material. You know that you're that much closer to getting an admission decision or scholarship. Regardless of how you did compared to other people, you know you did the best you could.
Break it into small, SMART goals.
SMART is an acronym that stands for Specific, Measurable, Achievable, and Relevant. Humans, especially those with mental health issues (which affect 1/4 of people), have a hard time achieving goals sometimes. We think of a goal as a huge task that has to be accomplished all at once. Instead, the best way to complete a goal is to split it up into pieces. For example: You want to make a 1500 on the SAT. You take the SAT the first time and score a 1300. You see that your weakest area was math. So you decide to study math. Don't say, "I'm going to master math by the time I take the SAT again", which leaves you with no direction. Instead, try saying, "I'm going to study one subsection of math every week for three months. Every day I'll study 30 minutes or complete 15 practice problems, whichever comes first." 30 minutes a day and one section a week is an attainable method of reaching your math goal.
Reward yourself intelligently upon achieving goals. By rewarding yourself every time you complete a small goal, you're wiring your brain to associate good feelings with your SAT prep. This reinforces your desire and motivation to study and will help you put in more hours without feeling bored. However, you must reward yourself after the goal is complete. This is especially true with addictive rewards- sugary foods, video games, drinking, TV, and technology. Sugar and alcohol (and some drugs) have a negative effect on cognition speed and memory. These rewards can sap your productivity. You also want to avoid settling into your favorite game or TV show after only 10 minutes of studying and realize hours later that you didn't finish all your homework, and you've still got more practice problems to do to meet your daily goal. Save the fun for afterward- you'll feel better and you'll be building better SAT prep habits.
Also, to achieve high SAT scores, keep in mind that the SAT is competitive, but don't get down on yourself. No matter what, don't beat yourself up.
The SAT is designed so that the results follow a Normal Distribution. This means that the scores fit a bell-shaped pattern, with 68% of students scoring between 400 and 600 per section.
You can see the score distribution for 2016 here.
Most of your SAT score comes from the preparation you received from your teachers. Most do not teach specifically SAT prep classes. Instead, they teach the component elements of the SAT test as school subjects. So, if your instruction was subpar, or you didn't do so well in a particular class, you won't have a good comprehension of that SAT subject. Also, the SAT test may ask questions that use familiar concepts in a new way. For example, one SAT question asks you to find the area of a shaded region inside a square with a circle (see picture).
Many geometry classes don't teach that. Also the SAT will use high-level vocabulary words in the reading section that students won't normally encounter, so they won't know the definitions.
Q: What's the minimum time I can study per day?
There is no minimum. It all depends on how much time you want to spend a week. However, it's important that you practice as close to everyday as possible. This facilitates maximum skill-building, as Daniel Coyle writes in his book The Talent Code. Here's the science behind it: the more frequently you do something, the more myelin wraps around the neurons in your brains. This makes increases the synaptic connections and makes the neural activity faster. It's better to spend 5 hours spread out over 5 days a week instead of 5 hours in 2 days. Higher frequency is better than longer sets.
You can also use Khan Academy to take a practice test that features unused questions written by the writers of the new SAT. Additionally, there are several study websites like PrepScholar that have SAT questions outside of a test, categorized by subject.
After you've taken the practice test and viewed your results, go back through the test and fill out the PowerScore Skills Assessment. It may seem like a lot of work, but trust us, this will actually boost your SAT score many times by identifying what your weakest areas are. Once you have identified your weaknesses, you can use any resource to help improve in those areas. The next few sections of this guide will focus on resources, courses and books for SAT prep.
Q: What tools do I need to study effectively?
You might find this list helpful.
Q: What are some highly effective study strategies I can use?
You can visit this link for more information.
Q: How do I choose the best study environment?
Noise, lighting, music, and distractions all play a role in deciding your study environment. For more information, you can read this guide.
If you don't have a dime to your name, don't worry- there are several resources you can use for free.
Students who first took the PSAT, then studied on Khan Academy for 20 hours and took the SAT scored 115 points higher on the SAT on average. Khan Academy offers practice tests, thousands of practice questions, and videos, all for free. It also can import your PSAT and tell you immediately what areas you should practice on. Overall, Khan Academy is your best free go-to resource for the breadth and quality of what it covers.
Union Test Prep offers practice tests, flashcards and study guides for the SAT. The flashcards are really just practice questions chosen at random for different subjects. The study guides are great, because they summarize an entire broad topic of math, reading or writing into a single guide.
This playlist of YouTube videos for the new SAT is a collection of tips and tricks that will make a big difference in your test-taking skills. Of course, to be most effective, you will need to also invest hours in studying the material, but these tricks will help you move faster and more efficiently through the test.
If you're looking for more than free content and have the money, then paid options are a good idea. Here are some of the best.
An established giant in the field of SAT prep, Kaplan offers the complete package. Practice tests, videos, one-on-one help, 18 hours of live instruction, and two SAT prep books.
For $299 you get the self-paced version, perfect for those with busy schedules. It features 140+ videos, 14 practice tests, customized lesson plans, online drills to practice skills, and 3 hours of on-demand help from an instructor.
For $1099 you get the classroom experience, with in-person instruction and lots of Q&A, along with everything in the lower price tier.
PrepScholar offers thousands of test practice questions written by people who scored in the top 1% of all test takers and even by some who got perfect scores on the SAT, along with graduates from top universities. It personalizes your learning by showing you exactly where to focus to best improve your score.
CurveBreakers is more of a dedicated tutoring service than an all-around prep course. You can get online tutoring worldwide or in-person tutoring if you live in New York. Nick LaPoma, owner and main tutor of CurveBreakers, is a veteran in the field of test prep. Also, CurveBreakers has success stories of students increasing their SAT score on the old SAT by 310 points.
If you want a hard-copy version of the 8 SAT practice tests offered for free online, you can go with The Official SAT Study Guide, 2018 Edition. You'll need a separate app to score your tests in the book, though.
An alternative is SAT Prep Plus 2018: 5 Practice Tests + Proven Strategies + Online (Kaplan Test Prep). The Online Resources are more practice questions to help practice your SAT skills.
To get more of a strategic view on the SAT, pick up a copy of the SAT Prep Black Book 2nd Edition. It explains in detail how the SAT is designed to trick students and yet has only one correct answer per question.
A great SAT Reading book is The Critical Reader 3rd Edition. It has a breakdown of the reading section categorized by skill, so you can selectively train your weakest skills.
Steve Warner writes a series of SAT Math books that are tailored to your ability level- he writes separate books for beginners, intermediate and advanced SAT math test takers. It's important in math to have a strong foundation and his books will help you to build one or enlarge it if you're already strong in math, but want to get even better. Here's his advanced book.
For SAT Writing, we recommend Erica Meltzer's grammar book The Ultimate Guide to SAT Grammar. You can get a set of five SAT Grammar and Writing practice tests with 44 questions each with RK Test Prep's SAT Grammar book.
For those who already have high scores, the only way to improve is by mastering the material. However, for low to medium scorers, tips and tricks can make improvements on the SAT. Here are some useful ones.
Since the SAT is a multiple choice test, there can only be one right answer. There will always be reasons why three answers are incorrect and only one is correct.
Keep an eye out for words indicating positive or negative connotations in the part of the passage that the question refers to. This can help to eliminate some choices, even if you don't know which remaining choice is correct. Strong adjectives can help to pin down the connotation of the word or phrase. Also, look for contrast words like "however", "rather than" or still".
Even though the SAT math section gives you formulas, memorizing them lets you access them quicker and gives you a greater understanding of how they work in context. By memorizing formulas, you'll be able to identify when and where to apply them.
If two answer choices both fit the rules of proper grammar, pick the one that says its message in fewer words.
Now that you've seen resources and how to study for the SAT, here's what you don't want to do.
Studying like mad in a few days leading up to the test is certainly better than not studying at all. But the science shows that the better way to master the material is slow but steady. A UCLA study found that the lack of sleep associated with cramming erased the benefits of studying material right before the test. With the SAT, like most other long-term goals, consistency is key.
You get a coffee and drink it down within minutes and begin studying. But you crave that caffeine rush and want another cup. One more won't hurt, right? Caffeine can help you stay awake and boost your speed on certain tasks as a stimulant. 400 mg, or about four cups of coffee, is safe for adults, but too much can cause unwanted side effects. The worst of it is anxiety– excessive caffeine use can cause physiological symptoms like jitters and nervousness, which can manifest mentally as anxiety about upcoming events like the SAT! Caffeine can also become addictive and hard to quit. So think hard about your use.
Another UCLA study found that excessive sugar sabotages learning and memory. Sugar may give you a high in the short term, making you feel happier or more engaged in what you're studying, but it damages the brain and can exacerbate Alzheimer's in the long term. Replace sugar and carbohydrate-rich foods with protein and healthy fat-rich foods such as nuts and fish. Protein helps the regulation of dopamine and serotonin, which are as important for learning as they are for your feelings and mental health.
The night before the SAT, arrange your schedule so that you get as close to nine hours of sleep as you can. The morning of the exam, be careful to eat a good breakfast.
Avoid juices- they do have some vitamins, but they lose the fiber that makes them healthy and retain the sugar.
On the exam day, before you take the exam, exercise. Even just walking for 20 minutes can boost your scores. You'll also feel better and more optimistic about the test.
If you have time, meditate. Of course, meditation confers more benefits to those who practice it over time, but even just a little bit can help to clear your mind before the high-stakes SAT.
You just took the SAT. Now what?
One option is to take it again, but only if you have time to study before the next exam. If you have no time left before your college applications are due, then it's time to say goodbye to SAT prep and hello to college applications. It's never too early to start on your college application essay.
If you didn't quite meet your goal score on the SAT, don't worry. College applications take into account not only your SAT score, ACT score and GPA, but also your volunteering experience, your extra-curricular activities and achievements, and your application essay. So there are plenty of ways to get competitive for colleges besides just the SAT.
Overall, learn to accept your result and don't be self-critical. Self-criticism can sometimes motivate us to improve, but overdone it hinders us by making us want to avoid activities that generate self-criticism. Instead, opt for self-compassion and self-care, where you accept that you are a good person inside no matter what mistakes you have made. You don't need to be a Buddhist monk to appreciate that simple philosophy.
Note: This guide also applies for the PSAT, which is almost the same as the SAT with a few small differences. Mainly, the PSAT is a little shorter and a tad bit easier than the SAT and is taken in the fall of 10th/ 11th grade. Also, good scores on the PSAT qualify students for National Merit Scholarships. Your PSAT score should be a good predictor of your SAT score.


The existence of cancer stem cells in hepatocellular carcinoma (HCC) has been verified by characterizing side population (SP) cells based on efflux of Hoechst 33342 dye from stem cells. Recent advances in microRNA (miRNA) biology have revealed that miRNAs play an important role in embryonic development and tumorigenesis. However, it is still unclear which miRNAs participate in the neoplastic transformation of liver cancer stem cells (LCSCs) during hepatocarcinogenesis.
To identify the unique set of miRNAs differentially regulated in LCSCs, we applied SP sorting to primary cultures of F344 rat HCC cancer cells treated with diethylnitrosamine (DEN) and normal syngenic fetal liver cells, and the stem-like characteristics of SP cells were verified through detecting expression of CD90.1, AFP and CK-7. Global miRNA expression profiles of two groups of SP cells were screened through microarray platform.
A total of 68 miRNAs, including miR-10b, miR-21, miR-470*, miR-34c-3p, and let-7i*, were identified as overexpressed in SP of HCC cells compared to fetal liver cells. Ten miRNAs were underexpressed, including miR-200a* and miR-148b*. These miRNAs were validated using stem-loop real-time reverse transcriptase polymerase chain reaction (RT-PCR).
Our results suggest that LCSCs may have a distinct miRNA expression fingerprint during hepatocarcinogenesis. Dissecting these relationships will provide a new understanding of the function of miRNA in the process of neoplastic transformation of LCSCs.
Cancer stem cells (CSCs) have been identified in hematopoietic malignancies and in solid tumors, including hepatocellular carcinoma (HCC) [1, 2]. The isolation and characterization of CSCs are usually based on the presence of known stem cell markers, i.e., CD133 in glioma  and CD44 and CD24 in breast cancer . However, for many tissues, specific molecular markers of somatic stem cells are still unclear. Therefore, attempts have been made to identify CSCs in solid tumors through isolation of side population (SP) cells based on the efflux of Hoechst 33342 dye; such efflux is a specific property of stem cells . The ability to isolate SP cells by cell sorting makes it possible to efficiently enrich both normal somatic stem cells and CSCs in vitro without the use of stem cell markers.
HCC is one of the most malignant tumors in existence. By using SP sorting, the existence of liver cancer stem cells in many established HCC cell lines has been verified [6–8]. However, few studies have focused on the isolation and characterization of SP cells isolated from primitive HCC cells. We conjectured that if normal hepatic stem cells (HSCs) and liver cancer stem cells (LCSCs) could be enriched through SP isolation, an in vitro model to determine whether HCC arises through the maturational arrest of HSCs could be developed.
MicroRNAs (miRNAs) are noncoding RNAs of 19 to 25 nucleotides in length that regulate gene expression by inducing translational inhibition and cleavage of their target mRNAs through base-pairing to partially or fully complementary sites . Studies using the Dicer gene knockout mouse model have demonstrated that miRNAs may be critical regulators of the organogenesis of embryonic stem cells (ESC) [10, 11]. Moreover, accumulated data suggest that dysregulation of miRNA occurs frequently in a variety of carcinomas, including those of the lung, colon, stomach, pancreas and liver . The dual effects of miRNAs in both carcinogenesis and differentiation of normal stem cells strongly suggest that miRNA may be involved in the transformation of normal stem cells into cancer stem cells. Therefore, screening for differences in miRNA expression between normal HSCs and LCSCs should help to elucidate the complex molecular mechanism of hepatocarcinogenesis.
In this study, we applied SP analysis and sorting to F344 rat HCC cells induced with DEN and to syngenic rat day 14 embryonic fetal liver cells. After isolation of total RNA, microarray analysis of miRNA expression was performed in order to detect possible differences in expression levels of specific miRNAs in the two side populations. We found that 68 miRNAs were over-expressed in the side population of cancer cells compared to that obtained from fetal liver cells, while 10 miRNAs were relatively under-expressed. Partially dysregulated miRNAs were validated by real-time PCR analysis. Our results reveal that miRNAs may play an important function during the transformation of normal HSCs into LCSCs.
Pregnant F344 rats and normal male F344 rats were purchased from the national rodent laboratory animal resources, Shanghai branch, China. All animals were housed in an air-conditioned room under specific pathogen-free (SPF) conditions at 22 ± 2°C and 55 ± 5% humidity with a 12 hour light/dark cycle. Food and tap water were available ad libitum. All operations were carried out under approval of Fourth Military Medical University Animal Ethics Committee. Primary HCCs were induced with DEN (80 mg/L in drinking water, Sigma, St. Louis, MO) for 6 weeks; animals were then provided with normal water until the appearance of typical tumor nodules in the liver, which usually occurred 10 to 12 weeks after treatment. After the rats were sacrificed under ether anesthesia, liver tissues were fixed with 4% paraformaldehyde, routinely processed and stained with hematoxylin and eosin (H&E) for histological examination by two pathologists, blinded to the results of the study, in order to verify the formation of HCC.
Fetal liver cells were obtained from embryonic day 14 rat fetuses by the procedure of Nierhoff et al. . The dissociated cells were inoculated onto culture plates with William's E medium (Sigma, St. Louis, MO) supplemented with 10% fetal calf serum (FCS) (Invitrogen), 100 U/mL penicillin G, 0.2 mg/mL streptomycin, and 500 ng/mL insulin. HCC cells were isolated from DEN-induced rat liver carcinomas. Briefly, tumor nodules in the liver were minced into pieces and digested by 0.5% collagenase type IV (Sigma, St. Louis, MO) at 37°C for 15 minutes. After filtration through 70 μm mesh, the dispersed cancer cells were collected by centrifugation and finally cultured in medium of the same composition as that used for fetal liver cells. The culture media were changed routinely every 3 days.
To identify and isolate SP fractions, fetal liver cells and HCC cells were dissociated from culture plates with trypsin and EDTA, and pelleted by centrifugation. The cells were resuspended at 1 × 106/mL in pre-warmed HBSS with 2% bovine serum albumin (BSA) and 10 mmol/L HEPES. Hoechst 33342 dye was added to a final concentration of 5 mg/mL in the presence or absence of 50 μM verapamil (Sigma, USA), and cells were then incubated at 37°C for 90 minutes. After incubation, the cells were washed with ice-cold HBSS three times, and were further stained with FITC-conjugated anti-rat CD90.1 monoclonal antibody (Biolegend Co., USA). When staining was finished, propidium iodide (PI; final concentration 1 μg/ml) was added to identify viable cells. The cells were filtered through 80 μm mesh (Becton Dickinson Co., USA) to obtain a single cell suspension before analysis and sorting. Analysis and sorting were performed on a FACSVantage II (Becton Dickinson Co., USA). The Hoechst 33342 dye was excited at 355 nm and its fluorescence was dual-wavelength analyzed with emission for Hoechst blue at 445 nm, and Hoechst red at 650 nm.
Total RNA from two groups of SP cells was isolated using TRIZOL reagent (Invitrogen) according to the instructions of the supplier and was further purified using an RNeasy mini kit (Qiagen, Valencia, CA USA). The miRCURY Hy3/Hy5 labeling kit (Exiqon) was used to label purified miRNA with Hy3TM fluorescent dye. Labeled samples were hybridized on the miRCURY LNA (locked nucleic acid) Array (v.11.0, Exiqon, Denmark). Each sample was run in quadruplicate. Labeling efficiency was evaluated by analyzing signals from control spike-in capture probes. LNA-modified capture probes corresponding to human, mouse, and rat mature sense miRNA sequences based on Sanger's miRBASE version 13.0 were spotted onto the slides. The hybridization was carried out according to the manufacturer's instructions; a 635 nm laser was used to scan the slide using the Agilent G2505B. Data were analyzed using Genepix Pro 6.0.
Signal intensities for each spot were calculated by subtracting local background (based on the median intensity of the area surrounding each spot) from total intensities. An average value of the three spot replicates of each miRNA was generated after data transformation (to convert any negative value to 0.01). Normalization was performed using a per-chip 50th percentile method that normalizes each chip on its median, allowing comparison among chips. In two class comparisons (embryonic hepatocytes SP vs. HCC SP), differentially expressed miRNAs were identified using the adjusted t-test procedure within the Significance Analysis of Microarrays (SAM). The SAM Excel plug-in used here calculated a score for each gene on the basis of the observed change in its expression relative to the standard deviation of all measurements. Because this was a multiple test, permutations were performed to calculate the false discovery rate (FDR) or q value. miRNAs with fold-changes greater than 2 or less than 0.5 were considered for further analysis. Hierarchical clustering was generated for both up-regulated and down-regulated genes and conditions using standard correlation as a measure of similarity.
To compare the expression of AFP and CK-7 between SP and non-SP and validate the differential expression of miRNAs in SP fractions, we applied real-time RT-PCR analysis to sorted cells. Specially, stem-loop primers were used for reverse transcription reaction of miRNAs . The complementary DNA (cDNA) underwent 40 rounds of amplification (Bio-Rad IQ5) as follows: 40 cycles of a 2-step PCR (95°C for 15 seconds, 60°C for 60 seconds) after initial denaturation (95°C for 10 minutes) with 2 μl of cDNA solution, 1× TaqMan SYBR Green Universal Mix PCR reaction buffer. The sequence of primers used for amplification is listed in Table 1. mRNA or miRNA levels were normalized using GAPDH or U6 RNA as a internal reference gene and compared with non-SP cells. The relative amount of each miRNA to U6 RNA was described using the 2-∆∆Ct method .
Cells sorted by FACS were washed twice with ice-cold PBS and then incubated with ice-cold cell lysis buffer (1% Nonidet P-40, 50 mmol/L HEPES, pH7.4, 150 mmol/L NaCl, 2 mmol/L ethylenediaminetetraacetic acid, 2 mmol/L phenylmethylsulfonyl fluoride, 1 mmol/L sodium vanadate, 1 mmol/L sodium fluoride, and 1× protease inhibitor mixture) to extract protein. The protein concentrations of the lysates were measured using a Bradford protein assay kit (Bio-Rad). All samples were separated in 12% SDS polyacrylamide gels. Signal were revealed by primary antibodies and IRDye700-labeled secondary antibody. The signal intensity was determined by Odyssey Infrared Imaging System (LI-COR Bioscience, Lincoln, NE).
The existence of the SP fraction in primary fetal liver cells and in HCC cells was confirmed by staining with Hoechst 33342 dye to generate a Hoechst blue-red profile. A small fraction of low-fluorescing cells in the lower-left region of each profile was gated as SP. The appearance of this fraction was blocked by verapamil, an inhibitor of transport via multidrug resistance proteins (Figure 1A-D). Both fetal liver cells and HCC cells contained a distinct fraction of SP cells. The SP of fetal liver cells was calculated to be 0.15% ± 0.02% (mean ± SEM), and that of HCC cells was calculated to be 0.20% ± 0.08%. Once identified, the cells in the SP gate were sorted into a centrifuge pipe by FACS.
SP cell and non-SP cells analysis. (A and C) Representative side populations (SP) were identified in the P3 gate on the flow cytometry profile after the cells were stained with Hoechst 33342, (B and D): The SP cells in both HCC cells and fetal liver cells disappeared (0.0%) when cells are treated with 50 μM verapamil. (E-H) Analysis of stem cell marker expression on the surfaces of SP and non-SP cells. The number within each histogram represents the percentage of CD90.1 positive cells. (I-K) Quantitative analysis of AFP and CK-7 genes expression applied to sorted SP cells and non-SP cells by using Real-time RT-PCR. Data were normalized by using GAPDH housekeeping gene as endogenous control. (* P < 0.05, ** P < 0.01). (L-M) Western-blotting analysis of AFP and CK-7 protein expression in SP cells and non-SP cells. The relative expressions of protein were calculated through comparing with GAPDH protein.
To examine whether SP cells are enriched for characteristics of stem cells compared to the non-SP cells, we further characterized the SP cells from the fetal liver cells and HCC cells by analyzing the presence of markers known to be expressed commonly on the surface of HSCs. FACS analysis showed that CD90.1 positive cells made up 45% ± 2.7% of total SP from fetal liver cells, and 37% ± 2.1% of total SP from HCC cells. In contrast, only 0.1% ± 0.0% (fetal liver cells) and 0.8% ± 0.1% (HCC cells) were CD90.1 positive cells in non-SP fractions (Figure 1E-H). We next quantitatively compared the expression of AFP and CK-7 genes between sorted SP cells and non-SP cells. Real-time RT-PCR analysis revealed that AFP and CK-7 mRNA level in SP from the fetal liver cells were increased 4.3-fold and 1.9-fold, respectively compared to non-SP (Figure 1I). Similarly, in SP from the HCC cells, they were increased 3.6-fold and 2.7-fold, respectively (Figure 1J). Furthermore, the differentially gene expressing profile of AFP and CK-7 in sorted SP cells and non-SP cells also confirmed by using western-blotting analysis. As shown in Figure, the relative expression of AFP and CK-7 were 0.84 ± 0.10, 0.53 ± 0.01 in SP from the fetal liver cells. While they were only 0.20 ± 0.08 and 0.18 ± 0.05 in non-SP cells (Figure 1L). Similar results also could be seen in HCC cells group (SP: 1.17 ± 0.0.14, 0.47 ± 0.10; non-SP: 0.35 ± 0.12, 0.16 ± 0.04) (Figure 1M). These results indicate that the SP fraction appeared to be enriched with HSCs or LCSCs.
To identify specific miRNAs that might function in neoplastic transformation of liver cancer stem cells, we analyzed global miRNA expression using miRCURY LNA Array that covered all microRNAs in miRBase. Slides were scanned using an Agilent G2565BA Microarray Scanner System and image analysis was carried out with ImaGene 7.0 software (BioDiscovery). The array data was further analyzed using SAM. Based on the fold-changes observed, 68 up-regulated miRNAs and 10 down-regulated miRNAs were identified in the SP of HCC cells compared to the fetal liver cells. A comprehensive list is shown in Table 2. The SAM analysis plot image is shown in Figure 2, and a hierarchical clustering image is shown in Figure 3.
SAM outputs. SAM plotsheet outputs under the four sets of criteria: Δ = 0.25, fold change = 2. Conditions are indicated at the upper right corner of each plotsheet. The red, green, and black dots represent upregulated, downregulated, and insignificantly changed miRNAs, respectively. The upper and lower 45° degree lines indicate the Δ threshold boundaries. The number of significant miRNAs, median number of false positives, and false discovery rate (FDR) are indicated at the upper left corner of the plotsheet.
Heat map of altered miRNA expression. A heat map was generated using the expression ratios of 78 miRNAs that differed significantly in SP of HCC cells compared to fetal liver cells, according to significance analysis of microarrays (SAM). Red, overexpressed miRNAs; green, underexpressed miRNAs compared to counterparts. Relatedness in miRNA expression across samples is shown by a hierarchical tree on the Y axis through standard linkage.
Using a stringent cut-off of P < 0.05, we found significantly altered expression of only 7 of all rat miRNAs analyzed in SP of HCC cells. In detail, five miRNAs were significantly up-regulated (miR-21, miR-34c-3p, miR-470*, miR-10b, let-7i*) and two miRNAs significantly down-regulated in SP of HCC cells (miR-200a*, miR-148b*). miRNA-specific qRT-PCR was used to validate the significantly altered miRNAs from the miRNA microarray results. As shown in Figure 4A, the results showed that the expression levels of miR-21, miR-34c-3p, miR-16, miR-10b, and let-7i* in SP of HCC cells compared to SP of fetal liver cells were increased 3.5 ± 0.84, 2.1 ± 0.52, 2.2 ± 0.46, 3.9 ± 0.61, and 2.8 ± 0.25 -fold respectively, which were consistent with miRNA microarray results (P < 0.05). of the down-regulated miR-200a*, and miR-148b* in SP of HCC cells had the fold changes 0.1 ± 0.04, and 0.4 ± 0.08, respectively (P < 0.01).
Validation of microarray data using real-time RT-PCR. (A) The levels of miR-21, miR-34c-3p, miR-470*, miR-10b and let-7i* are significantly increased, while the levels of miR-200a*, miR-148b are significantly decreased in the SP of HCC cells compared to the fetal liver cells, according to the results of microarray analysis (gray bar). Real-time RT-PCR analysis of these miRNAs using total RNA isolated from the SP fractions showed similar results (white bar). (B) Real-time analysis revealed that some known target genes of those partially validated miRNAs are also significantly differentially expressed between the SP sorted from the HCC cells and fetal liver cells (* P < 0.05; ** P < 0.01). The levels of target gene mRNA are inversely correlated with associated microRNA expression in SP cells.
To further confirm the differentially expressed miRNA, some known target genes expression of those validated miRNAs excluded miR-470* and miR-148b were detected in sorted SP cells and compared by using qRT-PCR between fetal liver cell and HCC cells. These target genes were PTEN (miR-21), P53 (miR-34c), Rho C (miR-10b), RAS (let-7i), and ZEB1 (miR-200a). As shown in Figure 4B, the relative gene expression of PTEN, P53, RhoC and RAS in SP from HCC cells were significantly lower than in fetal liver cells. On the contrary, the relative expression of ZEB1 gene in SP from HCC cells was higher than in fetal liver cells. Respectively, corresponding specific data were 0.78 ± 0.24 vs 0.33 ± 0.18 (PTEN), 1.79 ± 0.36 vs 0.81 ± 0.29 (P53), 1.16 ± 0.44 vs 0.72 ± 0.34 (RhoC), 3.52 ± 1.13 vs 1.62 ± 0.92 (RAS), and 0.27 ± 0.11 vs 0.48 ± 0.13 (ZEB1). These data were indirectly validated the differentially expressing profile of those miRNAs in SP fractions between HCC cells and fetal liver cells.
There is a growing realization that many cancers may harbor a small population of cancer stem cells (CSCs). These cells not only exhibit stem cell characteristics, but also, importantly, are tumor-initiating cells and are responsible for cellular heterogeneity of cancer due to aberrant differentiation. According to the hierarchical model of cancer, the origin of the cancer stem cells may be long-lived somatic stem cells. Therefore, markers of "normal" stem cells are being sought with the expectation that these molecules are also expressed by cancer stem cells, and can be used to identify them. In fact, the specific markers of many somatic stem cells, e.g., HSCs, are still unidentified, and it is difficult to screen putative stem cell markers useful for isolation and characterization of liver cancer stem cells. Recently, however, a special common "marker" has been identified in the sense that characteristic stem-like cells possess an energy-dependent drug export property conferred by their high expression of ABC (ATP-binding cassette) membrane transporters. This property was first exploited by Goodell et al.  for isolation and analysis of hematopoietic stem cells based on their ability to efflux a fluorescent dye. Identified cells were termed a "side population". The SP fraction is a useful tool for cancer stem cell studies in solid tumors, especially when specific cell surface markers are unknown. In many gastrointestinal cancers and HCC cell lines, SP fraction cells have been identified and characterized by their capacity for self-renewal and their high tumorgenicity . These studies demonstrated that SP can be used to enrich cancer stem cells in HCC. Moreover, it has been verified that normal HSCs (or 'oval cells') in rodents also express the side population phenotype defined by high expression of ABC transporter [18, 19]. In the current study, we were able to identify a small SP component (0.10%-0.34%) in both fetal liver cells and HCC cancer cells of F344 rats. The percentage of SP cells we detected is similar to the percentages described in most previous reports of SP in human HCC cell lines. To the best of our knowledge, this is the first report demonstrating the existence of SP cells in both fetal liver cells and in primary rodent HCC cancer cells induced by chemical carcinogens. Since the HCC cancer cells and fetal liver cells used in our study originated from the same inbred rat strain, the SP fractions enriched by screening both normal fetal liver and tumors for stem-like cell characteristics have high similarity in genetic background, thus providing a model for in vitro study of the mechanism of neoplastic transformation from normal HSCs into LCSCs. In contrast, it is difficult to accomplish this using SP cells sorted from many human HCC cell lines.
Increasing evidence has accumulated suggesting that many miRNAs play key roles in stem cell maintenance and differentiation. In ESC, disruption of the Dicer protein, an important enzyme in miRNA processing, leads to embryonic lethality . Further evidence has also been provided by studies in some somatic stem cells showing that specific miRNA-based regulation is involved during organ and tissue development; e.g., a cardiac-enriched miRNA family was identified and demonstrated to have a critical role in the differentiation and proliferation of cardiac progenitor cells . Additionally, experiments using isolated populations of hematopoietic stem cells have documented roles for specific miRNAs in HSC lineage differentiation, and evidence suggests that miRNAs are important for differentiation of somatic stem cells in several other tissues as well . In addition to stem cell studies, microarray-based expression studies have also shown that aberrant expression of miRNAs occurs in several hematological and solid tumors including HCC . In these malignancies, it has been shown that specific miRNAs can function either as oncogenes or as tumor suppressors during carcinogenesis . Moreover, the aberrant miRNA expression profile correlated with particular tumor phenotypes can even be used to distinguish between normal tissue and tumors.
With the accumulation of evidence for "cancer stem cells", it is proposed that miRNAs might play a role in malignant transformation from normal stem cells into cancer stem cells. Recent studies have partially verified this hypothesis; e.g., let-7 miRNA expression can be observed in ESC and progenitor cells, but is absent in breast cancer stem cells. The reintroduction of let-7 into these cells causes differentiation and reduction of proliferation and tumor-forming ability. It has been demonstrated that in carcinogenesis, some miRNAs are likely to be instrumental in helping to control the delicate balance between the extraordinary ability of stem cells to self-renew, and their ability to differentiate for the purpose of development and tissue maintenance versus their potential for dysregulated growth and tumor formation . In the present work, we have identified, for the first time, miRNA expression patterns that can unambiguously differentiate LCSCs and normal HSCs, though both were enriched in SP fractions and showed similar phenotypes. Our study demonstrates that the aberrant expression of some specific miRNAs may play a key regulatory role in the hepatocarcinogenesis of HSCs. Notably, the dysregulated miRNAs identified in our study are encoded in chromosomal regions that have frequent chromosomal instability during hepatocarcinogenesis, verified by previous comparative genomic hybridization. For example, the precursor sequences of the up-regulated miRNAs (miR-21, miR-10b) and down-regulated miR-148b* observed in our study are located at 17q23, 3q23 and 12q13. In these regions, chromosomal aberrations such as recurrent amplification, methylation or loss of heterozygosity have been detected in various clinicopathological HCC samples [25, 26]. It has been shown that miRNA expression profiles of cancer stem cells are tissue-specific and tumor-specific. Moreover, comprehensive analysis of miRNA expression in diverse tumors has shown that miRNA genetic fingerprints can be used to accurately diagnose and predict tumor behavior [27, 28]. While liver cancer stem cells are believed to be the tumor-initiating cells of HCC, we speculate that screening of circulating miRNAs in the serum could help to predict the presence of liver cancer stem cells and that such a procedure may be useful for early diagnosis of HCC.
Here we validated significant overexpression of miR-10b, miR-21, and miR-34c-3p in SP fractions of HCC compared to SP fractions of normal fetal liver cells. Notably, overexpression of these three miRNAs was previously shown to be an important factor in promoting cell invasion or proliferation in various tumor types. By performing real-time PCR, Sasayama et al.  found that miR-10b expression was upregulated in gliomas and that the expression of miR-10b was associated with higher-grade glioma. In glioma cells, miR-10b regulates the expression of mRNA for RhoC and urokinase-type plasminogen activator receptor (uPAR) via inhibition of translation of the mRNA encoding homeobox D 10 (HOXD 10), resulting in invasion and metastasis of glioma cells. Similarly, overexpression of miR-10b was also detected in metastatic breast cancer by Ma et al. , who showed that increased expression of miR-10b promoted cell migration and invasion. Additionally, it has been verified that miR-21 overexpression can down-regulate the Pdcd4 tumor suppressor and stimulate invasion, intravasation and metastasis in colorectal cancer . Moreover, overexpression of miR-21 was also previously associated with poorly differentiated HCC, and this miRNA is known to participate in down-regulation of phosphatase and tensin homolog (PTEN) . A different situation exists with other miRNAs such as miR-34c-3p, which is a member of the miR-34 family. Members of this family have been shown to be targets of the p53 gene, and to be involved in control of cell proliferation . However, since inactivation of p53 is a critical event during hepatocarcinogenesis, it has been suggested that miRNAs play a central role in the aberrance of the p53 tumor suppressor network during neoplastic transformation of liver cancer stem cells, and that this is linked with multiple changes of phenotype such as cell cycle arrest and apoptosis.
A subset of miRNAs was also identified and shown to be significantly underexpressed in our study, including miR-200a and miR-148b*. Previous studies have linked the miR-200 family with the epithelial phenotype , and Korpal et al.  identified miR-200a as a suppressor of epithelial-mesenchymal transition (EMT) through direct targeting of ZEB1 and ZEB2 genes. EMT is a crucial process in the formation of various tissues and organs during embryonic development. Moreover, EMT is proposed to be a key step in the metastasis of epithelial-derived tumors including HCC. Thus, we hypothesize that the down-regulated miRNAs seen in this study may function as tumor suppressor genes during carcinogenesis. Although the exact target mRNA targets for many miRNAs are currently unknown, use of the TargetScan and MiRanda database to identify predicted target genes of the miRNAs shown to be up-regulated or down-regulated in our study could help to elucidate the neoplastic mechanism of liver cancer stem cells.
This work provides an in vivo model for the study of mechanisms of neoplastic transformation of liver cancer stem cells by separately sorting SP fractions enriched with stem-like cells from primary rat HCC cancer cells and syngenic fetal liver cells. On the basis of this model, differences in miRNA expression profiles between LCSCs and normal HSCs were investigated using microarrays. This allowed us to identify miRNAs whose deregulation was closely correlated with the malignant phenotype of liver cancer stem cells, as distinguished from normal hepatic stem cells and from oncogene and tumor suppressor gene mutations. The gene and protein networks directly targeted and affected by these miRNAs that are likely to participate in tumorigenesis remain to be explored.
Ren Li, Niansong Qian, Kaishan Tao, Nan You contributed equally to this work.
This work was supported by grants from the National Natural Science Foundation of China (No. 30772102 and No. 30772094). We thank Professor Qinchuan Zhao for helpful suggestions in the preparation of the manuscript.
LR and DKF designed the study. LR performed cell isolation and cultures. QNS performed the western-blotting and analyzed the data statistically. TKS performed quantitative PCR analysis for target genes of validated miRNAs. YN performed miRNAs microarray detection and data analysis. WXC accomplished quantitative PCR validation. LR wrote the main manuscript. DKF looked over the manuscript. All authors read and approved the final manuscript.


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UMM leadership change announced
July 18, 2019 News
Andrew Egan, vice president for academic affairs and head of campus at the University of Maine at Machias (UMM), has announced his intention to step down, effective Aug. 15.
Egan joined UMM, a regional campus of the University of Maine, on Aug. 13, 2018.
Daniel Qualls, associate professor of education, will serve as interim vice president for academic affairs and head of campus. Qualls joined the UMM community in 2008 and holds a Ph.D. from the University of Tennessee.
"We appreciate Andy's leadership in the second year of the UMaine-UMM partnership and wish him well," says UMaine President Joan Ferrini-Mundy, to whom the UMM vice president and head of campus reports. "I look forward to collaborating with Dr. Qualls, the UMM Board of Visitors and the UMM community to sustain and grow the University of Maine at Machias as an educational, cultural and economic resource for the Down East region."
"It's been a real honor serving the UMM community and the hardworking people of Down East Maine," Egan says. "I wish UMM the very best going forward."
Promoting enrollment growth, increasing efficiencies, and advancing program and research opportunities are at the heart of the primary partnership between UMaine and the University of Maine at Machias that began July 1, 2017. At that time, UMM became a regional campus of UMaine, while maintaining its mission, degree programs and leadership role in the community.
Contact: Margaret Nagle, 207.581.3745
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