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Fac igitur calculum the legality versus ethics in vesica formatum ductum claudere, puttermesser papers quid accedit? One or two specimens have been already given,[106] but the reader may not regret the trouble of perusing the following in addition. De passione .
The legality versus ethics
Legality the versus ethics. [L] In the middle states also, many people pronounce a t at the end of once and twice , oncet and greek gods essay twicet . In point of fact, then, it is perfectly plain that we have only two parties in the field: Of these, four are aspirates, ef, es, eth, and esh. Furthermore, as touching that Praier, that nothing borne within the house might ever proove good, it is not haply meant of any persons, man or woman, but of dogges rather which were whelped there; which ought to be, not kinde and gentle, but curst and terrible. Or be misunderstood to assert, that a supposed revelation cannot be proved false, from internal characters. Such, briefly, was the political Dissertation on powerpoint stick needle injury condition of Spain, internally and externally, when news arrived of what is a letter of intent to purchase the occurrences at Nootka Sound which have been discussed in the foregoing chapters. Indeed, his heavy, pink-tea attentions were obviously regarded as quite flattering by the fair recipients! At 9 o'clock on the evening of May 6, the day of the debate in Parliament just studied, Miranda respect definition paper again met Pitt on the latter's invitation. the history of holocaust There are mark twain: thematic response Planche's extravaganzas, and later Dion Boucicault's versatilities—"classical" comedies like "London Assurance," sentimental Irish melodramas—"The Shaughraun," "The Colleen Bawn"—and popular favorites, such as "Rip Van Winkle"; the equally versatile Tom Taylor, with his "Our American Cousin," "The Ticket-of-Leave Man," etc.; Burnand's multifarious facetiae ; the cockney vulgarities of that very prolific Mr. Sheridan's plays, on the other hand, have never lost their popularity as acting dramas. Another had his thigh so tightly pressed that he felt the pain for a long time. Malone, whose opinion is supported by very solid reasons, thinks it was argumentative essay organ donation written in 1600; and therefore to reconcile the explanation given by Mr. I., they would be used simply to recommendation letter for mechanical engineering job illustrate or explain difficult passages in Shakespeare's text. As muscles are capable the transcendentalism in thoreau of shortening and elongating nearly a memoir personal essays fourth of their length, they readily produce the precise kind and degree of motion required in any particular case.[16] [16] The extensor muscles preponderate in mass and weight over the flexors, but this is readily accounted for by the fact, that the beard s thesis extensors, when limbs are to be straightened, always work at a mechanical disadvantage. Tabitha, after a faint resistance, falls into his humor and proves an the legality versus ethics the legality versus ethics apt pupil in the ways of worldliness. Multominus assertionem suam roborabunt, postquam intaminatum conspexere cadaver sanguine refertum, ut infra probabo. Paradoxical indeed is the attitude of mind of practically everyone toward this subject of being ushered out of life. As they are spread abroad in the air, they see in the clouds what must happen, and thus foretell the rain which they were aware of before it had been felt upon earth. Eccles. Neither onions, parsnips, carrots, nor cabbages are here. "The probabilities," says STEWART, "resulting from a concurrence of different analogies, may rise so high as the legality versus ethics to produce an effect on the belief scarcely distinguishable from moral certainty." When it is considered that strategic management and leadership in singapore airlines Butler's argument is wholly in addition to the cumulative mass of direct and almost irresistible evidence, and removes even the objections which attend the subject, we see the rejection of Christianity to be inexpressibly rash and absurd. Therefore he did what all should do--he acknowledged the hand of the Lord in all Example of a methodology section for a research paper things, in adversity as well as prosperity. Inst. It exists because of the efforts of hundreds of volunteers and donations from people in all walks of life. The the legality versus ethics speed of the bird as compared with that of the fish, or the speed of the fish as compared with that of the man, is no criterion of the power exerted. The former having conquered Troy, and having united themselves under one common name and interest, began, from that period, to distinguish the rest of the world by the title of barbarians ; inferring by such an appellation, "that they were men who were only noble in their own country; that they had no right, from their nature , to authority or command; that, on the contrary, so low were their capacities, they were destined by nature to obey , and to live in a state of perpetual drudgery and subjugation."[022] Conformable with this opinion was the legality versus ethics the treatment, which was accordingly prescribed to a barbarian . the legality versus ethics Those of Africa , situated near the line, of the same colour. 110:1-4, 11. All this while he made the hand and fore arm be secured in a cylinder of strong pasteboard, and the legality versus ethics suspended in a sling. In the way of waste ---- This expression is from the same law manufactory referred to by Mr. They are a sufficient answer to the charge, sometimes made, that the Latter-day Saints rank Joseph Smith as next in dignity and power to Jesus Christ. This is a perfectly legitimate line of argument, but before resorting to polyandry for an explanation of the Aryan levirate, it is worth while to inquire whether there is anything in known Aryan customs capable of supplying an the legality versus ethics explanation. He interposed in such a manner as was necessary and effectual to prevent that execution of justice upon sinners, which God had social sciencesffecaddcaddc appointed should otherwise have been executed upon them; how to prepare business plan or in such a manner, as to prevent that punishment from actually following, which, according to the legality versus ethics the general laws of divine government, must have followed the sins of the world, had it not been for such interposition.[214] If any thing here said should appear, upon first thought, inconsistent with divine goodness; a second, I am persuaded, will entirely remove that appearance. 327, in which, but not in the quotation from it, it appears that the rapier actually was in use in the time of Henry the Eighth; and therefore it is impossible to decide that this weapon, which, with its name, we received from the French, might not have been known as early examples of argument essay as the reign of Henry the Fourth, or even of Richard the Second. Let him then take in the consideration, of what importance religion is to mankind. By R. He has a few moments yet, and so he strolls over to a door at the opposite side of the building. This instance may be adduced with that of Philinium, and the young woman of the Rue St. The strawberries, the raspberries, the celery, the turnips, wave green above the clean earth, with no enemy in sight. [1] 2 Cor. It is likewise at times dangerous to extirpate these tumors, on account of their situation. And that the female has even the greatest share in the production of the foetus, will be proved by this instance: I almost think that, with one of these in the hands of an ordinary day-laborer, you might see at night where he had the legality versus ethics been working. F Ditto posterior margin. And is n't it better to buy little by little, enjoying every new object as you get it, and assimilating each article to your household life, and making the home a harmonious expression what is a proposal paper for college of your own taste, rather than to order things in sets, and turn the legality versus ethics your house, for the binge crinking is a problem time being, into a furniture ware-room? It follows from this that those insects and birds are endowed with the greatest powers of flight whose wings are the longest. [118] Hosea ii. And Tibullus says the legality versus ethics that he has seen the enchantress bring down the stars from heaven, and turn aside the thunderbolt the legality versus ethics ready to fall upon the earth--and that she has opened the ground and made master thesis routing protocols the dead come forth from their tombs.
This state of discipline may be necessary for the display of character.= Not to the all-knowing Being, but to his creation, or part of it, and in many ways which we know not.= It may be a means in disposing of men according to character.= the legality versus ethics And of showing creation that they are so disposed of.= Such display of character certainly contributes, largely, to the general course of things considered in the legality versus ethics this chapter. And objections from this head are not objections against revelation; but against the whole notion A short story of alfred bernhard nobel of religion, as a trial: For suppuration is the work of nature, an effort of the sound parts, by which they throw off whatever is stance on current politics vitiated and noxious; and it is the business of the essay on patriotism in english 100 words surgeon to assist this salutary operation, by removing, with his instruments, such parts as are intirely corrupted; but this ought to be done, at least as much as possible, without causing any discharge of blood[14]. He is sometimes dressed up in armour and set on to attack the legality versus ethics the knights of Cornwall, who are uniformly described as cowards. The pixies, it is the legality versus ethics traditionally averred, so delighted in this spot that they would carry their elfin babes thither, and unpleasant experience essay writing sing them to rest. It was no sword of territorial conquest, but that flaming blade of conscience and self-conviction which lightened between our first parents and their lost Eden,--that sword of the Spirit that searcheth all things,--which severs one by one the ties of passion, of interest, of self-pride, that bind Book sites the soul to earth,--whose implacable edge may divide a man from family, from friends, from whatever is nearest and dearest,--and which hovers before him creative writing in georgia like the air-drawn dagger of Macbeth, beckoning him, not to the legality versus ethics crime, but to the legitimate royalties of self-denial and self-sacrifice, to the freedom which is won only by surrender of the will. It existed in the American colonies before they became independent states; and notwithstanding the maxims of equality which have been adopted in their what acting and theater is several constitutions, it exists in most, if not all, of them, at this day, in the persons of macbeth the witches our free Negroes and mulattoes; whose civil incapacities are almost as numerous as the civil the legality versus ethics rights of our free citizens. The patterne of painful adventures &c. Just as good men as creating a good thesis fought at Marathon or Agincourt. THAISA. FOOTNOTES: A very constant feature of the old drama was the professional fool, jester, or kept clown, with his motley coat, truncheon, and cap and bells. It has been judiciously remarked by the legality versus ethics one of the commentators, that we are not to attend on these occasions merely to philosophical truth, but to consider what might have been the received or vulgar notions of the time: In the matter of technical construction he followed in a general way, intuitively, unconsciously for the most part, without caesars death: a conspiracy elaborate examination, the form the legality versus ethics of a short story as he was acquainted with it through his reading of stories. Divided either pinion in the direction of its long axis into three equal parts, the anterior nervures being contained in the anterior portion. [294] [It is remarked by DEAN FITZGERALD, that "It is not inconceivable that the Almighty should elizebeth proctors anger apply such a test of men's candor and fidelity, as should require them first to act upon a thing as true, before they were so fully satisfied of its truth as to leave no doubt remaining. And they may , in like manner, and perhaps, in some further providential manner, with respect to moral and religious subjects: And here, unless report mis-say, The grey-beard dwells; and on homework help online phschool this day Keeps yearly wassel, ever gay, With festive mirth and glee. As a consequence, the under surface of the wing is, as a rule, inclined upwards and forwards, even in flexion, so that it acts as a kite in extension and flexion, and during the up and down strokes. But as it is not impossible,[300] surely, that this alleged doubtfulness may be men's own fault; it deserves their most serious consideration, whether it be not so. Could there be a the legality versus ethics more glaring instance of "straining at a gnat and swallowing a camel"? I think the open fire does not kindle the imagination so much as it awakens the memory; one sees the past in its crumbling embers and ashy grayness, rather than the future. Thus Spenser, "What voice of damned ghost from Limbo lake Or guileful spright wandering in empty ayre, Sends to my doubtful eares these speeches rare?" Fairy Queen , book i. The slight but necessary emendation of and for that ascribed to Johnson, had already been made by Hanmer. Project Gutenberg volunteers the legality versus ethics and employees expend considerable effort to identify, do copyright research on, transcribe and proofread public domain works in creating the Project Gutenberg-tm collection. This, however, was, as they confess themselves, very seldom submitted to in ulcers of the legs; and was rather inserted to complete their treatises, than from a belief that the operation ought to be insisted on. 556, and Steevens's Shakspeare, vol. While preachers who held the divine right of kings made the churches of Paris ring with declamations in favor of democracy rather than submit to the heretic dog of a Bearnois,--much as our soi-disant Democrats have lately been preaching the divine right of slavery, and denouncing the heresies of the Declaration of Independence,--Henry bore both parties in hand till he was convinced that only one course of action could possibly combine his own interests and those of France. I must, therefore, take three sections to clothe it. [496] Vit. And by doing the legality versus ethics this he choice addiction a essay is disease or a proved that he despised the very masses whose sweet breaths he wooed, and had no faith in the system under which alone such a analysis pest of south essays africa one as he could have been able to climb so high. Four days after the death of Louis XVI the British Cabinet decided to authorize St. "I would like you to find out, if you can, what we do in the other world. The unfortunate Africans, terrified at these repeated depredations, fled in confusion from the coast, and sought, in the interiour parts of the country, a retreat from the persecution of their invaders. And courage never to submit or yield"; the courage which "bates no jot of heart or hope, but still bears up and steers right onward." There is nothing more bracing in English poetry than those passages in the sonnets, in "Paradise Lost" and in "Samson Agonistes" where Milton speaks of his blindness. From the first moment, he fell into the ways of the house and assumed a recognized position in the family,--I say recognized, because after he became known he was always inquired for by visitors, and in the letters to the other members a short review of the wealth of nations a book by adam smith of the family he always received a message. If then the superstition about the bean was borrowed by the Italians, it must have been root canal research paper borrowed in primitive times; and we must think that the belief reached the Italians at the same time as the cultivation of the bean itself spread from its original women, equal rights, and war (unknown) home. It is fun to see their swinging legs dangling over the tail of every wagon. Compare Ether 3:14, and Alma 11:38, 39. "Bold, as a hero,, as a virgin, mild." So in the following. Indeed, if by a myth we mean a tale told about gods or heroes, there are the legality versus ethics no Italian myths.[20] Myths attached to Greek loan-gods were borrowed with the gods the legality versus ethics from Greece. Versus ethics legality the.
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text:
Q: How to get the coordinates of the following point knowing two previous points and the distance to the following point I am wondering how to find the coordinates of the following point If I know two previous points and distance to the following point. For example:
-----P1------P2----d---P3-----

Data:

*

*P1: { x: 0, y: 1 }

*P2: { x: 2, y: 1 }

*d: 2
Searched:

*

*P3: x3?, y3?
For this example, the result will be P3: { x: 4, y: 1 }. This is a straightforward calculation. Things get complicated when P1 and P2 are in different quadrant's, the direction of following point is changing or the line is under some tilts angle(for this I do not have any tests yet).
All these possibilities can be described by below picture(excluding these with line under some tilted angle):

For now, I end up with such algorithm:
interface Coordinates {
    x: number,
    y: number,
}

function getCoordinatesOfFollowingPointInDistanceToLastPointOnLine (p1: Coordinates, p2: Coordinates,
    distance: number): Coordinates {
    const lineSlope = (p2.y - p1.y) / (p2.x - p1.x);
    const lineTiltAngle = Math.atan(lineSlope);
    const p3x = Math.abs(p1.x) + Math.abs(p2.x) > Math.abs(p1.x)
        ? p2.x + distance * Math.cos(lineTiltAngle)
        : p2.x - distance * Math.cos(lineTiltAngle);
    const p3y = Math.abs(p1.y) + Math.abs(p2.y) > Math.abs(p1.y)
        ? p2.y + distance * Math.sin(lineTiltAngle)
        : p2.y - distance * Math.sin(lineTiltAngle);

    return {
        x: Number(p3x.toFixed(3)),
        y: Number(p3y.toFixed(3))
    };
}

I have test my algorithm, and it doesn't work in some cases(when line is horizontal and following point is in -∞ direction). All tests are shown below:
  getCoordinatesOfFollowingPointInDistanceToLastPointOnLine
    Orientation Horizontal
      Following point in +∞ direction
        √ should calculate correctly position of P3 if P1 and P2 are in I quadrant (2 ms)
        √ should calculate correctly position of P3 if P1 and P2 are in II quadrant (1 ms)
        √ should calculate correctly position of P3 if P1 and P2 are in III quadrant
        √ should calculate correctly position of P3 if P1 and P2 are in IV quadrant (1 ms)
        √ should calculate correctly position of P3 if P1 in quadrant II and P2 in quadrant I (1 ms)
        √ should calculate correctly position of P3 if P1 in quadrant IV and P2 in quadrant III (4 ms)
      Following point in -∞ direction
        × should calculate correctly position of P3 if P1 and P2 are in I quadrant (4 ms)
        × should calculate correctly position of P3 if P1 and P2 are in II quadrant (1 ms)
        × should calculate correctly position of P3 if P1 and P2 are in III quadrant (1 ms)
        × should calculate correctly position of P3 if P1 and P2 are in IV quadrant
        × should calculate correctly position of P3 if P1 in quadrant I and P2 in quadrant II
        × should calculate correctly position of P3 if P1 in quadrant III and P2 in quadrant IV (1 ms)
    Orientation Vertical
      Following point in +∞ direction
        √ should calculate correctly position of P3 if P1 and P2 are in I quadrant
        √ should calculate correctly position of P3 if P1 and P2 are in II quadrant
        √ should calculate correctly position of P3 if P1 and P2 are in III quadrant (1 ms)
        √ should calculate correctly position of P3 if P1 and P2 are in IV quadrant
        √ should calculate correctly position of P3 if P1 in quadrant IV and P2 in quadrant I
        √ should calculate correctly position of P3 if P1 in quadrant III and P2 in quadrant II (1 ms)
      Following point in -∞ direction
        √ should calculate correctly position of P3 if P1 and P2 are in I quadrant (1 ms)
        √ should calculate correctly position of P3 if P1 and P2 are in II quadrant
        √ should calculate correctly position of P3 if P1 and P2 are in III quadrant
        √ should calculate correctly position of P3 if P1 and P2 are in IV quadrant
        √ should calculate correctly position of P3 if P1 in quadrant I and P2 in quadrant IV (1 ms)
        √ should calculate correctly position of P3 if P1 in quadrant II and P2 in quadrant III

Tests Code:
// Orientation Horizontal -> Following point in +∞ direction
test('should calculate correctly position of P3 if P1 and P2 are in I quadrant', () => {
    const p1: Coordinates = { x: 0, y: 1 };
    const p2: Coordinates = { x: 2, y: 1 };
    const p3 = getCoordinatesOfFollowingPointInDistanceToLastPointOnLine(p1, p2, 2);

    expect(p3.x).toBe(4);
    expect(p3.y).toBe(1);
});
// Orientation Horizontal -> Following point in -∞ direction
test('should calculate correctly position of P3 if P1 and P2 are in I quadrant', () => {
    const p1: Coordinates = { x: 2, y: 1 };
    const p2: Coordinates = { x: 1, y: 1 };
    const p3 = getCoordinatesOfFollowingPointInDistanceToLastPointOnLine(p1, p2, 2);

    expect(p3.x).toBe(-1);
    expect(p3.y).toBe(1);
});
// Orientation Vertical -> Following point in +∞ direction
test('should calculate correctly position of P3 if P1 and P2 are in I quadrant', () => {
    const p1: Coordinates = { x: 1, y: 1 };
    const p2: Coordinates = { x: 1, y: 2 };
    const p3 = getCoordinatesOfFollowingPointInDistanceToLastPointOnLine(p1, p2, 2);

    expect(p3.x).toBe(1);
    expect(p3.y).toBe(4);
});
// Orientation Vertical -> Following point in +∞ direction
test('should calculate correctly position of P3 if P1 and P2 are in I quadrant', () => {
    const p1: Coordinates = { x: 1, y: 2 };
    const p2: Coordinates = { x: 1, y: 1 };
    const p3 = getCoordinatesOfFollowingPointInDistanceToLastPointOnLine(p1, p2, 2);

    expect(p3.x).toBe(1);
    expect(p3.y).toBe(-1);
});

I could add one if to detect this case but what if the line is under some tilt angle, then this will be very hard to decide how to calculate coordinates of the following point. So, the question is: How my algorithm should look like to make it work always?
Update 1
I have tried the algorithm linked by @Andreas How to find a third point given both (2 points on a line) and (distance from third point to first point), but it doesn't work for me unless I am doing something wrong. Below you can find how I implemented this:
function getCoordinatesOfFollowingPointInDistanceToLastPointOnLine (p1: Coordinates, p2: Coordinates,
    distance: number): Coordinates {
    const p1P2Distance = Math.sqrt((p2.x - p1.x) ** 2 + (p2.y - p1.y) ** 2);
    const xDirectionLength = (p2.x - p1.x) / p1P2Distance;
    const yDirectionLength = (p2.y - p1.y) / p1P2Distance;
    const p3x = p1.x + distance * xDirectionLength;
    const p3y = p1.y + distance * yDirectionLength;

    return {
        x: Number(p3x.toFixed(3)),
        y: Number(p3y.toFixed(3))
    };
}

A: The formula from the question I've mentioned in my comment works.
Your update calculates P3 from P1 (p1.x + ..., p1.y + ...) but it should be from P2
I've replaced your tests and objects with arrays, but the formula is the same.
I had to replace ** with the "old" Math.pow() because otherwise the fourth test would have failed because (p2.y - p1.y) ** 2 resulted in a NaN.
(~~ is used to get integers instead of floats)

const testCases = [
  /* P1, P2, distance, P3 */
    [[0, 1], [2, 1], 2, [ 4,  1]],
    [[2, 1], [1, 1], 2, [-1,  1]],
    [[1, 1], [1, 2], 2, [ 1,  4]],
    [[1, 2], [1, 1], 2, [ 1, -1]]
];

function getP3(p1, p2, distance) {
    const p1p2d = Math.sqrt(Math.pow(p2[0] - p1[0], 2) + Math.pow(p2[1] - p1[1], 2));
    const dx = (p2[0] - p1[0]) / p1p2d;
    const dy = (p2[1] - p1[1]) / p1p2d;
    const p3x = p2[0] + distance * dx;
    const p3y = p2[1] + distance * dy;

    return [~~p3x, ~~p3y];
}

testCases.forEach(({0: p1, 1: p2, 2: d, 3: e}) => {
  const p3 = getP3(p1, p2, d);
  console.log(p3, e, p3[0] == e[0], p3[1] == e[1]);
})

redpajama_set_name: RedPajamaStackExchange


text:
Q: Violación de segmento punteros C tengo un problema con "violación de segmento" en el código de más abajo. Más preciso es, cuando el código llega a la parte del if(comparacion_estacion()){} me da error en cierta iteración del while, cuando la lista llega a cierto string "US Universidad de Santiago" (la lista tiene almacenado las estaciones de metro: [Abreviacion] [Nombre estacion]).
La función comparacion_estacion sí sirve, osea, sí me retorna true (primera iteración) y false (siguientes) pero no se cual será el detalle que hace dejar el código a mitad.
Anexo:
si comento toda la parte del if comparacion_estacion(lista->nombre,combinaciones[i].inicio)){}, funciona bien el for y el while, osea, me recorre el struct y la lista,
Nodo *conectar_linea(Nodo *lista){
 Nodo *aux;
 aux = lista;
   for(int i=0;i<22;i++){
    while(lista != NULL){

        if(comparacion_estaciones(lista->nombre,combinaciones[i].inicio)){
            cout<<"entro al if"<<endl;

           if(comparacion_estaciones(combinaciones[i].linea_termino,"linea1 ")){

                lista=conectar_estacion(lista,linea1,combinaciones[i].inicio,combinaciones[i].termino);
            }

            if(comparacion_estaciones(combinaciones[i].linea_termino,"Linea2 ")){}

            if(comparacion_linea(combinaciones[i].linea_termino,"Linea5 ")){
                cout<<"Exito"<<endl;
            }else
                cout<<"No son iguales";

            if(comparacion_estaciones(combinaciones[i].linea_termino,"Linea4 ")){}

            if(comparacion_estaciones(combinaciones[i].linea_termino,"Linea4a ")){}

            if(comparacion_estaciones(combinaciones[i].linea_termino,"Linea6 ")){}

        }

        else{
            puts("No se pudo conectar estaciones");
        }
        cout<<lista->nombre<<endl;
      lista=lista->sig;
      cout<<lista->nombre<<endl;
    }
    lista = aux;
}
return lista;
}

Adjunto también función comparacion_estaciones
    bool comparacion_estaciones(string estacion1, string estacion2 ){
       char modificar1[5], modificar2[4];
       char *token1, *token2;
strcpy(modificar1,estacion1.c_str());strcpy(modificar2,estacion2.c_str());
token1 = strtok(modificar1," ");token2 = strtok(modificar2," ");

  if(*token1 == *token2) {

    return true;}
else{
    return false;
}
}

Cuando ejecuto el programa
     ./programa
    entro al if
    Exito
    SP San Pablo
    NP Neptuno
    No se pudo conectar estaciones
    NP Neptuno
    PJ Pajaritos
    No se pudo conectar estaciones
    PJ Pajaritos
    LR Las Rejas
    No se pudo conectar estaciones
    LR Las Rejas
    EC Ecuador
    No se pudo conectar estaciones
    EC Ecuador
    AH San Alberto Hurtado
    No se pudo conectar estaciones
    Violación de segmento

La información de la lista:
    SP San Pablo
    NP Neptuno
    PJ Pajaritos
    LR Las Rejas
    EC Ecuador
    AH San Alberto Hurtado
    US Universidad de Santiago
    EL Estación Central
    LA ULA
    RP República
    LH Los Héroes
    LM La Moneda
    CH Universidad de Chile
    SL Santa Lucía
    UC Universidad Católica
    BA Baquedano
    SA Salvador
    MM Manuel Montt
    PV Pedro de Valdivia
    LE Los Leones
    TB Tobalaba
    GO El Golf
    AL Alcántara
    EM Escuela Militar
    MQ Manquehue
    HM Hernando de Magallanes
    LD Los Dominicos

A: El problema es dificil de entender porque falta más de la mitad del código. En el programa tienes varias funciones que manipulan y gestionan la lista y sus nodos y solo has puesto dos de ellas.
Aun así ya te puedo adelantar que una lista no es un nodo, de la misma moneda que una rueda no es un coche. Para no repetir, puedes ver más información al respecto en esta otra respuesta.
Por otro lado, no veo motivo que justifique pasar de std::string a char*. La clase std::string nació precisamente para evitar los malos vicios de char*, tipo de dato donde es tremendamente sencillo meter la pata, y para muestra un botón:
bool comparacion_estaciones(string estacion1, string estacion2 ){
  char modificar1[5], modificar2[4];
  char *token1, *token2;

  strcpy(modificar1,estacion1.c_str());strcpy(modificar2,estacion2.c_str());
  token1 = strtok(modificar1," ");token2 = strtok(modificar2," ");

  if(*token1 == *token2) {
    return true;
  }else{
    return false;
  }
}

¿Por qué modificar1 y modificar2 tienen tamaños distintos? Tenemos que tener en cuenta esto a la hora de decidir qué estación ponemos la primera y cual la segunda? Si desbordamos cualquiera de los dos arrays el programa podría empezar a comportarse de forma errática...
Y por si esto no fuera poco, resulta que la función desborda siempre. ¿Cómo lo se si no puedo probar el código? Por los ejemplos que has puesto:
SP San Pablo
   NP Neptuno
   PJ Pajaritos
En base al código entiendo que estacion1 y estacion2 almacenan una línea completa del ejemplo anterior, es decir, almacenarán unos 10-15 caracteres... y resulta que copias ese chorizo en un array con capacidad para 4 caracteres (caso de modificar1) y para 3 caracteres (modificar2).
Con lo sencillo que es copiar un std::string en otro y, además, así te quitas de problemas:
bool comparacion_estaciones(string estacion1, string estacion2 ){
  std::string modificar1 = estacion1;
  std::string modificar2 = estacion2;

  char *token1 = strtok(modificar1.c_str()," ");
  char *token2 = strtok(modificar2.c_str()," ");

Además, piensa que los char* no se pueden comparar así como así. La línea
if(*token1 == *token2) {

No comprueba que el contenido de ambas cadenas sea idéntico... sino que comprueba si ambos punteros almacenan la misma dirección de memoria... teniendo en cuenta que las cadenas las estás copiando en modificar1 y modificar2 veo dificil que ambos punteros acaben direccionando la misma posición de memoria.
std::string posee una característica bastante versátil... iteradores. Los iteradores son objetos que nos permiten iterar (qué nombres mas curiosos usan en C++) los elementos que componen el objeto principal. En este caso con los iteradores podemos movernos a través de la cadena de caracteres. La ventaja de los iteradores es que se pueden hacer multitud de operaciones con ellos.
Así, por ejemplo, lo que a ti te interesa es el código de la estación, que casualmente ocupa siempre los dos primeros caracteres de la cadena. Puedes entonces quedarte con ese rango y construir una nueva cadena a partir de el:
std::string modificar1(estacion1.begin(),estacion1.begin()+2);
std::string modificar2(estacion2.begin(),estacion2.begin()+2);

O, si prefieres usar la STL en todo su esplendor:
std::string modificar1(estacion1.begin(),std::next(estacion1.begin(),2));
std::string modificar2(estacion2.begin(),std::next(estacion2.begin(),2));

Y ya está, ahora en modificar1 y modificar2 se almacena única y exclusivamente el código de la estación. Y como además estas variables son de tipo std::string, podemos usar el operador de comparación directamente:
if( modificar1 == modificar2 )
  return true;
else
  return false;

Claro que esta comparación se puede simplificar muchísimo:
return (modificar1 == modificar2);


Thanks very much for using Team Snap. We will post short "how to" tutorials on this website and under the Posts tab in Team Snap.
Following is an overview of the different ways to send messages in Team Snap. For the best results using Team Snap, please accept "notifications" (under the "more" tab in My Preferences.
Available in the app format only. It's not available in the web/browser format of Team Snap.
We will use Posts for larger messages that don't require real-time dispersal.
The benefit to using a Team Snap email is that the recipient receives an alert to indicate an email is waiting to be read.
We will use Alerts to send urgent messages, e.g., ride cancellations or messages that you want the person to receive right away, e.g., ride changes, availability changes, etc.
For technical questions, please contact D'Arcy at 705-772-8425 or through Team Snap.


text:
12/3/2019 - Common Council Unanimously Adopts 2020 Budget
City of Kingston Common Council
Unanimously Adopts 2020 Budget
KINGSTON, NY -- Mayor Steve Noble's 2020 Recommended Budget was unanimously adopted by Kingston's Common Council tonight. The total 2020 General Fund budget is $44,464,333.
Under the 2020 Adopted Budget, the tax levy will remain frozen at $17,650,940 with a 0% increase for the fifth year in a row. Tax rates continue to decline for the third consecutive year for homestead (from $9.73 to $9.04 per $1,000 of assessed value), and a steep tax rate drop for non-homestead, which will decrease from $15.59 to $14.24 per $1,000 of assessed value. Over $1,070,000 has been placed in contingency in preparation for potential non-recurring expenses, keeps all essential services intact with no eliminations of currently filled full-time positions, and adds or expands 12 full-time positions.
The Common Council voted to appropriate an additional $542,000 from the fund balance to purchase equipment and vehicles for various City departments rather than bonding, which will save future interest costs for taxpayers.
"I want to thank the Common Council for their close read of the 2020 Budget and their diligence in looking at every facet when allocating taxpayer dollars. We have created a very strong budget that aligns with my overall goals for the future of the City of Kingston," said Mayor Noble. "The budget also introduces an incremental raise of $5000 a year for the next five years, which will bring the Mayoral salary to $100,000 in 2024. The last raise was in 2006 - this increase is commensurate with similar nearby cities and will ensure that the position of Mayor in the City of Kingston remains attractive to talented candidates with municipal experience."
"A $44,464,333 budget is a big deal. It's a lot of money and it's the lifeblood of the City. I want to assure the public that this budget has gone through extensive review. We held a series of meetings and every line item has been reviewed down to the last detail," said Finance Committee Chair Douglas Koop, Ward 2.
The 2020 Adopted Budget will support a variety of core service areas within the City of Kingston, including:
Infrastructure improvements continue to be a primary focus, in several areas: sidewalks, streets, waterfront, and sewer. Many infrastructure improvements and capital projects will be moving forward in 2020, including the Broadway Streetscape Project, Rondout Riverport Stabilization, Henry Street Safe Routes to School, Franklin Street Complete Streets, among others. The budget also includes essential DPW and Parks & Recreation equipment, including a dump truck, milling attachment for paving work, and a sweeper.
The Kingston City Land Bank will be fully funded in 2020, with a two-year combined investment of over $3 million into quality housing that is affordable. The Office of Economic and Community Development will move to a refurbished property on Franklin Street, ensuring that the Office's services are easily accessible to local residents.
The City will invest in necessary safety gear for the Kingston Police Department, including ballistics vests and other protective equipment. The Kingston Fire Department will purchase a new fire engine.
In 2020, the Parks and Recreation Department will continue to manage 110 acres of parkland, including 11 parks, 3 community centers, a riverfront beach, a community pool, and an interpretive nature center, as well as taking on management of Dietz Stadium. The budget earmarks $150,000 for Wi-Fi in Kingston parks, improvements to Block and Loughran Parks, as well as the construction of a skatepark at Hasbrouck Park.
Kingston continues to be a leader in environmental and sustainability initiatives. In 2020, the City will implement LED streetlight replacement, invest in several projects that protect and revitalized the waterfront, and continue to reduce emissions with electric vehicles and charging stations.
The 2020 Adopted Budget will be posted at: www.kingston-ny.gov under Latest News.

redpajama_set_name: RedPajamaCommonCrawl


text:
Slavery's long afterlife in the north
The anti-Blackness that thrives in the shadow of America's more blatant record
Policing Black Lives: State Violence in Canada From Slavery to the Present
Robyn Maynard
292 pages, softcover
At an event for her book at Montreal's Grande Bibliothèque late last year, Robyn Maynard asked the audience of about five hundred to raise their hands if they had been taught in school about the two hundred years of Black enslavement in Canada. One man raised his hand. One out of five hundred. This in a city, that, along with Quebec City and Halifax, was one of the transatlantic network ports, that frequently received ships containing enslaved Black men and women arriving from the Caribbean. A city where, as Maynard recounts in her methodically researched book Policing Black Lives: State Violence in Canada From Slavery to the Present, enslaved Black women were "particularly fashionable and noteworthy possessions." (She is quoting from Charmaine A. Nelson's Slavery, Geography and Empire in Nineteenth-Century Marine Landscapes of Montreal and Jamaica.) In 1734, one such enslaved Black woman, Marie-Joseph Angélique, was arrested, tortured, and publicly hanged after she was accused of burning down a large portion of Montreal as she fled her white mistress.
"To be Black in Canada is to live in slavery's 'afterlife,' " Maynard writes in the introduction to a book that painstakingly shows how an anti-Black mindset that was perfected in order to justify the ownership of human beings continues to taint and poison almost every aspect of Black life in Canada today.
Canada's willful amnesia to its history of anti-Blackness began early. Slavery in Canada was abolished in 1834. Just 31 years later, "textbooks bore little allusion to any Black presence in Canada, erased two centuries of slavery, included no mention of segregated schools (an ongoing practice at the time) and alluded to the issue of racial discord only in the United States," Maynard writes.
Being the neighbour of a behemoth can be a blessing and a curse. Canada might resent its neighbour's geopolitical heft and narcissism, but the United States' more blatant racist record has proven a useful foil that ensures "anti-Blackness…continues to hide in plain sight" north of the border, she argues. "When acknowledged, [anti-Blackness] is assumed to exist, perhaps, but in another time (centuries ago), or in another place (the United States)," writes Maynard.
Had Maynard asked her audience in Montreal for the names of victims of anti-Black police violence in the U.S., undoubtedly many more than just one would have been able to recall the names of Trayvon Martin and Michael Brown—and perhaps more—while at the same time being oblivious to the fate of Quilem Registre, a 39-year-old Montreal man who died in 2007 after police shocked him several times with a Taser, or Andrew Loku, 45, who was shot dead by a Toronto cop in 2015. Maynard is determined not just to honour those unknown and erased victims in Canada through witness and information, but to demolish the "wall of silence" that she says insulates state violence against Black persons in Canada.
The Canada that she knows "in the eyes of many of its citizens, as well as those living elsewhere, is imagined as a beacon of tolerance and diversity," is the same Canada where, Maynard notes, there are "enormous racial inequities with respect to income, housing, child welfare rates, access to quality education and health care and the application of drug laws."
Maynard places her book resolutely in that gap between what Canada likes to think it is and what it actually is, and makes it impossible to nurture the "fiction" that the state is the protector of its national subjects, as many believe. "It is more accurate to say that the state protects some at the expense of others," she contends. And she has numbers and facts that will ensure you cannot look away:
– Despite comprising 3.5 percent of the Canadian population, African Canadians are incarcerated in federal prisons at more than double that rate.
– Because so much of Canada's Black population was born elsewhere, significant numbers of those eventually released will be punished again by deportation.
– Black migrants are disproportionately affected by punitive immigration policies such as immigration detention and deportation, which, Maynard argues, is in part due to the heightened surveillance of Black migrant communities.
– Black children and youth in Canada's largest city are vastly overrepresented in state and foster care (41 percent in care are Black, compared with just 8.2 percent of the underage population who are Black) while they are far more likely to be expelled from high school. In a recent public school report in Toronto, nearly half of those expelled were Black.
– Black communities are, after Indigenous communities, among the poorest racial groups in Canada.
As Maynard makes clear, many of those facts have been documented by various Black Canadian scholars whom she readily acknowledges as inspirations and mentors. Yet, Canada remains known, locally and internationally, "as the land of multiculturalism and relative racial harmony."
Maynard is clear from the outset that her book's writ is to explore and expose anti-Blackness in Canada but she is insistent that we see historical and current connections between Black and Indigenous oppression. Her language is clear and brooks no subterfuge, or polite denial: "any study of institutionalized anti-Blackness must necessarily be informed by the reality that Canada is a settler colony founded on colonization and genocide," she writes.
In settler colonies, Maynard explains, Indigenous people are seen as being "in the way," therefore state violence comes in the forms of laws and policies aimed at destroying Indigenous communities to secure "unfettered access to Indigenous land." "Contrastingly, in the logic of Black enslavement, it is Black personhood that is under attack: 'the slave' is a useful commodity but 'the person underneath is imprisonable, punishable, and murderable.' " As Maynard notes, "the living legacy of slavery and the ongoing practice of settler colonialism at times result in similar forms of repression" that are obvious in Canada today: "Black and Indigenous peoples experience grossly disproportionate incarceration, susceptibility to police violence, poverty, and targeted child welfare removal."
If Canadians know the names of victims of anti-Black police brutality in the U.S. but not here at home, it is instructive that they also remember just the names of men.
Given that most Canadians are oblivious to the anti-Blackness that Maynard carefully documents, it is also unlikely they would approach Canada's bloody legacy with Indigenous peoples with the clear-eyed insistence on justice that Policing Black Lives demands of us. As the CBC reported last year, even after the launch of a national inquiry and an increased focus on the issue, there is still no way to tell how many Indigenous women and girls go missing in Canada each year. The inquiry, expected to cost $53.8 million, has already lost two executive directors and several staff members. All this at a time when Indigenous women represent just two percent of Canada's population, yet account for more than 25 percent of female murder victims in recent years.
In an era in which optics dominate, a conventionally attractive Canadian prime minister is regarded a champion of all the downtrodden because, for example, he wore "Eid Mubarak" socks to Toronto Pride. The bar is, after all, set exceptionally low: The elected leader of Canada's neighour to the south is an accused sexual predator and racist and Islamophobe. Maynard's book unflinchingly demands that we see past those optics. Right-wing groups have co-opted the phrase "social justice warrior" and turned it into an epithet against anyone who challenges fascism, or racism, or misogyny. But as I read Policing Black Lives, I understood Maynard was the most vital and focused definition of a social justice warrior.
Two of the book's most important contributions are possible precisely because its author is a Black feminist activist whose social justice work lives and breathes among the communities she writes about. Firstly, with regards to state violence, Maynard tells us that this is a concept too often equated with police brutality. But she insists on an expanded definition that also encompasses violence administered by other institutions outside of the criminal justice system, including institutions that many regard as administrative, such as immigration and child welfare departments, social services, schools, and medical institutions.
Secondly, Maynard writes, state violence and state-sanctioned violence are not evenly distributed among populations, but instead "deeply infused along the lines of race, class, and gender." Just as state violence is not just about police brutality, it is not always directed at just Black men, as is often imagined. "All Black people are not demonized equally or identically," Maynard writes. "Gender, sexual orientation, (dis)ability, mental health, and place of birth also mediate the way that anti-Blackness is experienced."
If most Canadians know just the names of victims of anti-Black police brutality in the U.S., it is instructive that they—like many Americans—remember just the names of men. Where are the women? It is obvious from Maynard's chapter on law-enforcement violence against women that many Black women are punished for being "uppity," refusing to embody submissiveness or deference to white law-enforcement officers. Take Majiza Philip, a Montreal woman charged with assaulting two police officers and obstructing justice outside a concert. (She was charged after knocking on the window of a police car, and acquitted in court last year.) Or Sharon Abbott, a newspaper delivery worker in Toronto who was followed by a white officer in 2007 as she delivered papers at three in the morning. The officer claimed he was following her because she was driving erratically and not wearing a seat belt. Abbott alleged that the officer violently subdued her and kept her handcuffed in his car for an hour. She was issued seven tickets for minor traffic violations; of the six that proceeded to a justice of the peace, Abbott was acquitted on all charges. Abbott, who sustained several injuries during her arrest, successfully filed a human rights complaint and won a $5,000 settlement. The tribunal found that Sergeant Stephen Ruffino's actions were "consistent with a manifestation of racism whereby a white person in a position of authority has an expectation of docility and compliance from a racialized person and imposes harsh consequences if that docility and compliance is not provided."
Maynard notes that the case of Chevranna Abdi, a 26-year-old Black transgender woman taken into police custody in 2003, was a reminder "that Black transgender women often live at the intersection of both the societal demonization of Black women and a societal hostility towards transgender persons." Law enforcement officers in Hamilton, Ontario, arrested Abdi then handcuffed her and dragged her, face down, down seven flights of stairs. She died soon after.
In each of her book's chapters Maynard pays particular attention to Black women, sexual minorities across the gender spectrum, and people with disabilities and-or mental illnesses. Determined to centre "historically expendable lives," Maynard writes that it is her intention "that this work also challenges widely held dichotomies of who is innocent-guilty and how this affects our perception of who is undeserving-deserving of state violence or protection."
To that end, she emphasizes the need for a recalibration of how we see all members of the Black community, regardless of where on the spectrum of "good-bad" they are placed by the law or "Euro-Christian morality." (The two are sometimes inseparable.) Whether "community members stigmatized by real or imagined involvement in illicit economies, those youth or adults who have used or sold illegal drugs, those whose gender presentations or sexualities have been deemed deviant, sex workers, undocumented migrants, and incarcerated persons," these Black persons, Maynard says, are "stigmatized and rendered disposable," leaving them the "most vulnerable to state abuse, exploitation, confinement, and even death with little or no outcry."
Similarly, Maynard clearly details the criminalization of ways in which poor Black women survive their poverty, and the accompanying accusations of "welfare fraud." She reminds us of the slavery-era stereotyping and demonization of Black women as "jezebels" who were sexually insatiable—"unrapable" is Maynard's chilling description— and whose mere presence in public space indicated a criminal sexuality. The echoes of that today are found in the ways that many Black cis and trans women are profiled and prosecuted as sex workers.
In that regard Policing Black Lives joins revolutionary texts by Black women scholars and feminists that are helping to unpack centuries of toxic and dangerous intersections of anti-Blackness and misogyny, such as the groundbreaking study Say Her Name: Resisting Police Brutality Against Black Women by Kimberlé Crenshaw and Andrea J. Ritchie, and to popularize the use of the word "misogynoir," coined by queer Black feminist Moya Bailey to identify the various forms of hatred directed at Black women.
Maynard is a fierce advocate. And a fierce optimist. "Being uniquely positioned at the intersections of multiple forms of societal and state violence has allowed Black women to be at the forefront of community-based anti-racist, feminist, queer-friendly, and class conscious responses to gendered and state violence in Canada," she says.
What would such responses be like? It is testament to Maynard's tenacious determination that she ends a book that focuses on oppression, violence, pain, and death with an affirmation of life and liberatory principles. Our world can be better, she says, and means it. Our imagination is bigger and better than borders, prisons, policing, and state surveillance, she tells us. "It is a matter, that is, of prioritizing life over death."
Mona Eltahawy is a feminist writer and public speaker who lives in Cairo and New York City. She is the author of Headscarves and Hymens: Why the Middle East Needs a Sexual Revolution.
A History of Silence
A review of Dying from Improvement: Inquests into Indigenous Deaths in Custody by Sherene Razack.
Stevie Cameron
The Gates of La Francophonie
A review of A Place in the Sun: Haiti, Haitians and the Remaking of Quebec by Sean Mills.
A Dangerous Isolation
A review of Police in Canada: The Real Story, by John Sewell
Susan Eng
Safer, Meaner Streets
A review of Canadian Policing in the 21st Century: A Frontline Officer on Challenges and Changes, by Robert Chrismas
Neil Boyd


Kansas running back Pooka Williams Jr. (1) is finally dragged down by the Rutgers defense after a long run during the first quarter on Saturday, Sept. 15, 2018 at Memorial Stadium.
The Kansas football team's overall struggles this season haven't stopped some talented individuals from standing out.
Both freshman running back Pooka Williams and redshirt senior linebacker Joe Dineen landed this week on national midseason All-America teams.
The Jayhawks' star back, Williams was named to ESPN's mid-year freshman All-America team. Dineen made an even bigger splash, as a first-team defender on Sports Illustrated's midseason All-America team.
Baylor quarterback Charlie Brewer (12) is tackled by Kansas linebacker Joe Dineen Jr. (29) during the first half of an NCAA college football game, Saturday, Sept. 22, 2018, in Waco, Texas.
The Jayhawks also were well represented on the midseason All-Big 12 teams from Pro Football Focus.
With a season grade of 85.8, Williams was the lone KU offensive player among PFF's all-league choices.
The defensive team, though, featured not only Dineen (88.1 overall grade), but also redshirt senior defensive lineman Daniel Wise (82.3) and junior safety Bryce Torneden (90.1).
KU football coach David Beaty said Tuesday sophomore quarterback Miles Kendrick, who hasn't played since suffering a shoulder injury in Week 4 at Baylor, is available for this week's game at Texas Tech. The backup QB also was considered available at West Virginia, before KU's bye week.
Beaty said junior cornerback Kyle Mayberry, who was available in an "emergency" situation at WVU, and junior safety Mike Lee, who missed the loss to the Mountaineers, are back to being available, as well.
Asked whether junior receiver Evan Fairs, who has played in only four games, was injured or was taking a redshirt, Beaty said the 6-foot-3 wideout is injured.
"Well, he's not available, so he's injured," Beaty added.
Sophomore receiver Quan Hampton, KU's head coach also revealed, could be a game-time decision at Texas Tech (2:30 p.m. kickoff, FOX Sports 1). Hampton missed KU's home win over Rutgers and its road loss at WVU.


This 32 page booklet is part of the Educational Practices Series developed by the International Academy of Education. It focuses on effective mathematics teaching. Drawing on a wide range of research, it describes the kinds of pedagogical approaches that engage learners and lead to desirable outcome.


Chagas disease is most often transmitted to humans by Trypanosoma cruzi infected triatomine bugs, and remains a significant cause of morbidity and mortality in Central and South America. Control of Chagas disease has relied mainly on vector eradication. However, development of insect resistance has prompted us to develop a paratransgenic strategy to control vectorial transmission of T. cruzi. Here, the potential role of recombinant endoglucanases as anti-trypanosomal agents for paratransgenic application is examined. The surface of T. cruzi is covered by a thick coat of mucin-like glycoproteins that have been proposed to play a role in the binding of T. cruzi to the membrane surface of the vector gut. We hypothesize that disruption of these glycoconjugates could arrest parasite development in the vector and abort the transmission cycle. In this work, we examine the effects of recombinant Arthrobacter luteus β-1, 3-glucanase expressed via Rhodococcus rhodnii on T. cruzi Sylvio II strain.
The coding sequence for β-1, 3-glucanase was cloned in-frame to a heterologous promoter/signal sequence from the Mycobacterium kansasii alpha antigen gene resident in an E. coli/R. rhodnii shuttle vector. The resulting construct was confirmed by sequencing, and electroporated into R. rhodnii. Expression products from positive clones were purified from log phase cultures followed by dialysis into physiological buffers. Lysates and media were quantitated by ELISA against rabbit antibody specific to β-1,3-glucanase. Glucanase-positive samples were applied to live T. cruzi parasites in culture and viability accessed by spectrophotometric and fluorescent microscopic measurements. R. rhodnii-expressed β-1,3-glucanase exhibited toxicity against T. cruzi compared to controls when applied at 5 and 10% of the total culture volume. The decrease in cell viability ranged from a maximum of 50% for the media treatments to 80% for the filtered lysates.
These results suggest that recombinant β-glucanase could be a powerful addition to the arsenal of effector molecules for paratransgenic control of Chagas disease. In future studies, the ability of β-glucanase to function in combination with other effector molecules will be explored. Dual targeting of T. cruzi should not only slow resistance but also permit synergistic or additive lethal effects on T. cruzi.
Chagas disease, a parasitic disease endemic to South and Central America, is caused by the protozoan Trypanosoma cruzi and transmitted to humans via the feces of triatomine bugs. It affects approximately 8–11 million people, results in 50,000 deaths and accounts for the loss of 500,000 disability-adjusted life-years annually . Cases of Chagas disease have expanded globally, and are now reported in the Americas, Europe, Australia and Asia due to emigration of infected individuals from endemic parts of the world .
Chagas disease manifests in three distinct phases - acute, indeterminate and chronic. Infected individuals present in an initial acute phase that develops into a chronic phase in 20-40% of the patients. The chronic phase is characterized by the development of lesions in the nervous tissue of the heart, intestine and esophagus that result in progressive organ enlargement with potentially fatal complications. Patients who do not progress to the chronic phase of the disease exist in an indeterminate phase . The two drugs used worldwide in the treatment of Chagas disease, benznidazole and nifurtimox, are effective only against the acute stage of the disease . These treatments are marred by their many side effects including bone marrow toxicity, neural toxicity and severe nausea and vomiting . Moreover, strains of T. cruzi that are resistant to both drugs have emerged and resistance to one drug is usually coupled with resistance to the other .
Paratransgenesis is an alternative approach under development to reduce transmission of vector-borne diseases by eliminating carriage of pathogens by their arthropod vector hosts . The strategy focuses on understanding the microbial flora of the insect host at the developmental locus of the pathogen and genetically transforming selected bacteria to produce molecules that disrupt the target pathogen. Thus the bacteria act as a carrier or delivery agent – a Trojan horse – for the anti-pathogen molecules . We developed the paratransgenic strategy to control Chagas disease transmission by triatomine bugs over 10 years ago . In this strategy, the relationship between an important vector, Rhodnius prolixus, and its gut symbiont, Rhodococcus rhodnii, was exploited to disrupt the transmission of T. cruzi. R. rhodnii is a nocardiform actinomycete that aids R. prolixus with vitamin metabolism after its blood meal . These actinomycetes, obtained through coprophagy, are essential for the survival of the triatomine bugs. Aposymbiotic nymphs of R. prolixus do not reach sexual maturity . Because of its symbiotic association and proximity to the infective trypomastigote form of T. cruzi in the insect hindgut, R. rhodnii serves as an ideal organism for application in paratransgenesis . We initially demonstrated that genetically engineered R. rhodnii that expressed an anti-trypanosomal peptide, cecropin A, when introduced into R. prolixus, resulted in elimination or significant reduction of T. cruzi in experimentally infected bugs . More recent in vitro studies have shown that several anti-microbial peptides (AMPs) are even more effective at targeting T. cruzi when used in combination . This opens up the possibility of using multiple molecules synergistically to eliminate vector carriage of the parasite.
In this study, we investigate the potential role of endoglucanases as anti-trypanosomal agents. The surface of T. cruzi is covered by a thick coat of mucin-like glycoproteins. Many of these glycoproteins are developmentally regulated and have been proposed to play a role in the binding of the cell body and the flagellum of T. cruzi to the membrane surface of the vector gut, an integral step in T. cruzi maturation . We hypothesize that disruption of the glycoconjugates by endoglucanases could arrest parasite development in the vector and abort the transmission cycle. Arthrobacter luteus lyticase is a complex endoglucanase consisting of β-1,3-glucanase and alkaline protease that degrades β-1,3 and 1–6 glycosidic linkages . We previously demonstrated that purified lyticase from Arthrobacter was highly effective at lysing T. cruzi in vitro. Here we report that recombinant Arthrobacter luteus β-1,3-glucanase expressed via R. rhodnii effectively kills T. cruzi Sylvio II strain, suggesting that this molecule may be a powerful addition to the arsenal of effector molecules for paratransgenic control of Chagas disease.
The cDNA of β-1,3-glucanase from Oerskovia xanthineolytica (also known as Arthrobacter luteus strain 73–14) was a gift from Dr. Steve Slilaty . To facilitate subsequent cloning steps the 1.7 kb β-1,3-glucanase cDNA was excised from pOP95-15 by AgeI and SmaI digestion, and subcloned into the XmaI and EcoRV sites of pBlueScript KS, generating pBS-glucanase.
The plasmid pRrExpA is an E. coli/R. rhodnii shuttle vector. Protein expression from this plasmid is under the control of a heterologous promoter-signal peptide complex derived from the alpha antigen gene (MKα) of Mycobacterium kansasii. A modified pBlueSript SK MCS, lacking the EcoRV through KpnI sites, is located just downstream of this promoter. The β-1,3-glucanase cDNA was ligated in frame into the BamHI/SmaI sites of pRrExpA following excision from pBS-glucanase by HindIII digestion, Klenow fill-in and subsequent BamHI digestion, to generate pRr-glucanase. Transformants were selected on Luria-Bertani (LB) plates containing 50 μg/ml of carbenicillin (CAR). Plasmid DNA isolated by Qiagen columns from the clones was sequenced to verify insertion.
R. rhodnii electro-competent cells were generated by growing the cells in 500 mls of modified BHI (BHI with 0.1% Tween-80 and 2% glycine) at 30°C until mid-log phase. Cells were chilled on ice and harvested by centrifugation. The pellets were washed three times with a cold filtered-sterilized solution of 10% glycerol, 0.05% Tween-80, and resuspended in 1 ml of cold 10% glycerol after the final wash. These electrocompetent cells were stored as 100 μl aliquots at -80°C until ready for use.
For electroporation, competent cells were thawed on ice and mixed with 200 μg of pRr-glucanase DNA. 80 μl of this mixture were transferred to a chilled electroporation cuvette (Bio-Rad, 0.1 cm gap) and electroporated twice using a field strength of 18.5 kV/cm with a time constant of 5 ms on a BioRad Gene Pulser II. Samples were placed on ice between electrical pulses. Following electroporation, cells were allowed to recover in 1 ml of BHI at room temperature for 2 hours. 200 μl aliquots were spread onto BHI plates containing 25 μg/ml CAR. Plates were incubated at 28°C, and checked every 2 days for growth for up to 1 month.
Plasmid specific primers, Mkα5' (5' CAG AGC TGA GCG GGA AGA TTC 3') and OriR3' (5' GCA CGA CCA CAG CAA TAC 3'), were used for colony PCR screening of putative R. rhodnii transformants. Cycling conditions were an initial denaturation step for 3 mins at 95°C, followed by 30 cycles at 92°C for 30 sec, 54°C for 30 sec, and 72°C for 1 min, followed by final extension at 72°C for 3 min. PCR products were analyzed by electrophoresis on 1% agarose gels.
Rhodococcus specific 16S rDNA sequencing was performed to confirm the identity of each transformant. Genomic DNA (gDNA), isolated from each R. rhodnii transformant (MasterPure Gram Positive DNA isolation kit - Epicentre), was amplified using 16s-RrF (5' CTG GGT CTA ATA CTG GAT ATG 3') and 16s-RrR (5' TGC CAT TAC TAG CGA CTC 3') primers. Cycling conditions were 3 mins of denaturation at 95°C, followed by 30 cycles of 95°C for 30 sec, 52°C for 30 sec and 72°C for 1.5 min, followed by a final extension for 3 mins at 72°C. Amplification was confirmed by running a portion of the reaction mixture on a 1% agarose gel. Samples that amplified were subsequently purified from the reaction using the Qiagen PCR Purification kit per manufacturer's instructions, and sequenced using the PCR primers described above with the BigDye Terminator Reaction Cycle Sequencing Kit (Applied Biosystems). Sequences were then submitted for BLAST analyses at the GenBank database.
Cells from 100 ml mid-log cultures of transformed and untransformed (negative control) R. rhodnii were pelleted by centrifugation and the spent media frozen at -20°C. The pelleted cells were suspended in 10 ml of complex lysis buffer (100 mM Tris, 500 mM NaCl, 0.5 mM EDTA, 0.1% Triton X-100, 0.1% Tween-20, 8% glycerol, 250 mM urea, 5 mM 2-mercaptoethanol, 100 μg/ml PMSF, 1X HALT Protease Inhibitor Cocktail, 50 μg/ml lysozyme and 1U benzonase) and lysed by alternating the suspended cells between cooling on ice and vortexing at ten-minute intervals. Following complete lysis after 30 mins, cellular debris was removed by centrifugation at 15,000 rpm for 30 mins. Half of the lysate was flash-frozen and stored at -80°C. The remainder was dialyzed for 48 hours at 4°C into 1X TBS (25 mM Tris, 0.5 M NaCl, pH = 7.2) with Sigma protease inhibitors using a 2,000 MWCO Slide-A-Lyser dialysis cassette (Pierce), with one change of dialysis buffer at 24 hours. Dialysate was aliquoted, flash-frozen and stored at -80°C.
A rabbit polyclonal antibody to the CHEPGTQFRGRVDGD epitope of β-glucanase was produced by GenScript for this laboratory. The specificity and affinity of this affinity-purified antibody was determined via ELISA against Arthrobacter lyticase complex and the synthetic peptide used to produce the primary antibody. Five-log dilutions of lyticase (100 U/μl to 0.01 U/μl) or β-glucanase peptide (0.01 nM to 100 nM) were bound overnight at 4°C to 96-well plates. Wells were washed and blocked with 2.5% BSA in TBS-T (25 mM Tris, 0.5 M NaCl, pH = 7.2, 0.5% Tween-20), and the primary antibody was added at a 1:5000 dilution in TBS-T. After a 2 hour incubation at 37°C, the primary antibody was removed, wells washed as before and a 1:5000 dilution of anti-rabbit IgG HRP added. Plates were incubated at 37°C in the dark for one hour before being washed 3 times prior to detection with single component TMB (BioRad). Plates were then read at OD450 on a Molecular Devices SpectraMax M2 Fluorescent microplate reader. These experiments were repeated in triplicate.
Lysates and spent media from transformed and control R. rhodnii were separated using SDS-PAGE on 8% Tris-glycine gels (Invitrogen). Proteins were subsequently transferred to 0.2 μm pure nitrocellulose membranes. Subsequent to blocking with 5% non-fat dried milk in TBS-T, a 1:5000 dilution of the anti-glucanase antibody was added and the membrane incubated overnight at 4°C. Following three washes, a secondary anti-rabbit IgG coupled to HRP was added to the membrane at a 1:10,000 dilution for 1 hour at room temperature. Blots were developed by incubation in TMB.
Samples from transformed and control R. rhodnii diluted in TBS-T were added to triplicate wells on microtiter plates, and incubated for 2 hours at 37°C. After washing the wells in TBS-T, 2.5% BSA was added to block for 1 hour at 37°C. The primary antibody (GenScript USA), diluted to 1:5000 (in TBS-T), was removed after overnight incubation at 4°C. The secondary antibody used in these experiments was an anti-rabbit IgG coupled to HRP. 100 μl of a 1:5000 dilution of this antibody was added to each well, and allowed to incubate at 37°C for an hour in the dark. Detection of expressed recombinant protein was accomplished after 3 final washes by development in TMB. Plates were read at 450 nm on the Molecular Devices SpectraMax M2 Fluorescent Microplate Reader.
The toxicity of recombinant β glucanase produced by R. rhodnii in cell lysates and spent media against epimastigote cultures of T. cruzi Silvio II (ATCC #50823) was determined by a combination of spectrophotometric turbidity and live/dead fluorescent microscopy assays as described in Fieck et al. . Briefly, lysates from β glucanase recombinants were diluted in 1X PBS and added to triplicate wells of sterile tissue culture certified 96-well plates. 105 parasites were added to each well and the plates incubated at 28°C for a total of 96 hours. The optical density of the cultures was measured at 600 nm every 24 hours. After the incubation period, parasites from triplicate wells were pooled and stained for 1 hour with 1 μl of 5 mM Calcein-AM (Molecular Probes). After washing in 1.0 ml 1X PBS, brightfield and green fluorescent images were generated on a Nikon Eclipse 80i microscope. In these experiments, lysates and spent media from untransformed R. rhodnii were used as negative controls. No human subjects or animals were used in this study.
Sequence analysis revealed that the β-1,3-glucanase cDNA was successfully cloned downstream of the MKα promoter-signal peptide of the pRrExpA shuttle vector. This clone, pRr-glucanase, was subsequently electroporated into R. rhodnii. The transformation efficiency of actinomycetes was in the range of one transformant per 108 cells. A total of 40 putative R. rhodnii-glucanase transformants were selected after 3 days of growth on BHI plates containing 25 μg/ml CAR. Of these, four were randomly selected for further characterization. The remaining cells were propagated in BHI containing 25 μg/ml CAR and cryopreserved in 10% glycerol for future use.
Amplification of the 16S rDNA using Rhodococcus specific primers on isolated genomic DNA confirmed that the selected transformants were, indeed, R. rhodnii. A 1.4kb PCR product was obtained from two of the four selected clones. Sequencing of the PCR product confirmed that the recombinant organism was R. rhodnii. To confirm that the two remaining Rhodococcus transformants harbored pRr-glucanase, colony PCR was performed using plasmid specific primers that flank the insert. Only one of the four transformants yielded a 1.7 kb fragment that is indicative of the insert.
After a thorough and unsuccessful search for commercially available antibodies against β-glucanase, a synthetic antibody was specifically raised for the detection of this recombinant molecule. The affinity of this antibody was initially tested by examining its ability to bind commercially available Arthrobacter lyticase. Figure 1 shows that a 1:5000 dilution of this antibody is capable of detecting Arthrobacter lyticase at a limit of 0.005 units/μl. The antibody was synthesized against an epitope on the β-glucanase peptide that is part of the Arthrobacter lyticase complex. Therefore, synthetic β-glucanase peptide was used to generate a standard curve that could be used to determine the expression level of glucanase from transformed R. rhodnii (Figure 2). The antibody detected β-glucanase at concentrations as low as 10nM. Resolution of concentrations lower than 10nM was impossible from the present ELISA. The OD readings corresponding to these standards curves were used to calculate the concentration of recombinant β-glucanase from the R. rhodnii clones.
Binding affinity of polyclonal anti-glucanase antibody. The binding affinity of the polyclonal anti-glucanase antibody at a dilution of 1:5000 was determined against five-log dilutions of Arthrobacter lyticase. Note that this polyclonal antibody can detect β-glucanase in as little as 0.05 U/μl of Arthrobacter lyticase. Each point represents the mean with standard error bars of triplicate samples. Experiments were performed at least twice.
ELISA standard curve for β-glucanase. Decreasing concentrations of synthetic β-glucanase were titrated against 1:5000 dilution of the polyclonal anti-glucanase antibody to generate ELISA standard curves such as the one shown here. These curves were used to quantify recombinant β-glucanase expression from each R. rhodnii clone. All points represent the mean with standard error bars of triplicate samples. Experiments were performed at least twice.
After standardization, lysates and spent media from cultures of the R. rhodnii transformant were analyzed via ELISA to confirm expression of recombinant β-glucanase. The concentrations of recombinant β-glucanase were extrapolated from the standard curve of the synthetic peptide. ELISA detected β-glucanase at a lower threshold of approximately 100 nM from 0.5X spent media and 80 nM from cellular lysates (Figure 3). Interestingly, 0.5X dilution of the spent media and lysates revealed a higher concentration of recombinant peptide than undiluted samples (Figure 3). We conducted another set of experiments to explain this finding. (1) To remove the effect of the lysis buffer, samples were dialyzed twice. Double dialysis led to loss of the recombinant protein (Figure 3) and was not a viable method of purification; (2) ELISA revealed that non-specific binding of the dilution buffer by the antibody was not a factor (data not shown); (3) ELISA using synthetic β-glucanase peptide added to wild-type R. rhodnii cell extracts showed that cell extracts block the signal from synthetic β-glucanase peptide (data not shown). These experiments suggest that the higher signal observed in diluted samples of the cell lysate is due, in part, to dilution of the cellular extract, which enabled detection of the recombinant peptide. Since the spent media samples were not affected by cellular extracts, the increase in signal in these samples after dilution was due to the prozone or hook effect observed in many immuno-ligand binding assays .
Expression of β-glucanase from R. rhodnii Clone 1. Dialyzed lysates and spent media from transformed R. rhodnii was examined for expression of β-glucanase. Expression of β-glucanase was consistently higher in each of the diluted samples, suggesting that there is self-blocking at high protein expression levels. As expected, the expressed protein is found predominantly in the spent media. Bars represents the mean with standard error bars of triplicate samples wells.
Arthrobacter lyticase exerted approximately 50 percent growth inhibition of T. cruzi at the lowest concentration applied (1 U/μl) and complete inhibition at 10 and 100 U/μl (Figure 4), demonstrating that lyticase is lethal to T. cruzi at very low concentrations. Recombinant peptide samples were therefore diluted to 5% and 10% for toxicity studies. As seen in Figure 5, recombinant β-glucanase isolated from filtered spent media inhibited T. cruzi growth by 50% at both dilutions. Filtered lysates containing recombinant β-glucanase were also diluted to 5% and 10%. These samples inhibited T. cruzi growth by ~80% without any significant difference in the growth inhibition between the two dilutions (Figure 5). Lyticase buffer and lysates from untransformed R. rhodnii (negative controls) showed 0% inhibition of T. cruzi growth (Figures 4 and 5).
Arthrobacter lyticase inhibited the growth of T. cruzi Sylvio. T. cruzi Sylvio was cultured in media with indicated concentrations of Arthrobacter lyticase. Parasite turbidity was determined following 96-hour incubation and compared to untreated parasites. Results were averaged from triplicate samples in two independent experiments, and are presented as the percent change in turbidity between treated and untreated cultures following the 96-hour incubation.
T. cruzi Sylvio is inhibited by recombinant β-glucanase. Indicated amounts of dialyzed lysates or spent media, containing recombinant β-glucanase, harvested from transformed R. rhodnii were added to cultures of T. cruzi Sylvio. Turbidity of parasites treated with lysates from untransformed R. rhodnii (control lysate) was compared to that of the treatment group after 96-hours. Results are presented as the percent change in turbidity between treated and untreated cultures following this incubation period. The result shown here is a representative from two independent experiments.
Microscopy studies confirmed the results we observed from spectrometric toxicity assays. Brightfield and fluorescence microscopy demonstrated a significant reduction in the number of live T. cruzi cells after they were treated with recombinant β-glucanase. Figure 6 shows the microscopy images of T. cruzi treated with lysates isolated from untransformed R. rhodnii alongside T. cruzi treated with lysates from β-glucanase-expressing R. rhodnii. Bright field images show a marked decrease in number of T. cruzi after treatment with recombinant β-glucanase. Most of the cellular debris seen under the bright field after treatment do not fluoresce nor have the structural integrity of healthy T. cruzi suggesting that they are lysed, unviable remnants of T. cruzi. Additionally, since only live cells take up the fluorescent Calcein AM and we see a decrease in fluorescent bodies after treatment, we can conclude that the treatment is effective in lysing T. cruzi.
Microscopic analysis of T. cruzi Sylvio following treatment with recombinant β-glucanase. T. cruzi Sylvio were treated with either cell lysates from untransformed R. rhodnii, or dialyzed lysates containing recombinant β-glucanase. Following 96-hour treatment, the parasites were harvested, stained with Calcein-AM and examined at 40X.
β-glucanase is a stable molecule with cruzicidal activity and, therefore, may play a part in a paratransgenic strategy for the control of Chagas disease. Given the critical role of the glycocalyx of T. cruzi in parasite development within the arthropod gut, we believe that disruption of the sugar coat using this molecule will abort vectorial transmission. Here we report the development of transformed R. rhodnii that express recombinant β-glucanase. This was confirmed by the results from ELISA of lysates and spent media isolated from an engineered line of this symbiont. Our results suggest that the cellular lysates contain a higher concentration of recombinant β-glucanase than surrounding media. We observed that the blocking effect of the cellular extract combined with a marked prozone phenomenon resulted in under-estimation of the concentration of recombinant β-glucanase in cell lysates. Nevertheless, toxicity assays revealed that any effect of T. cruzi inhibition can be attributed entirely to the recombinant β-glucanase. In ongoing studies, we are developing expression systems for R. rhodnii that permit enhanced secretion of heterologous proteins. We expect these constructs to improve the efficacy of β-glucanase as an effector molecule in the gut of triatomine bugs. However, high rates of bacterial turnover in the gut of these bugs, especially after blood meals, will liberate recombinant molecules as well [15, 24].
Development of recombinant β-glucanase as an effector for paratransgenesis adds another molecule to our growing arsenal. In vitro studies have already established that AMPs used in combination have synergistic lethal effects against T. cruzi. β-glucanase could be a powerful addition to the system when used in combination with other molecules. Drug resistance of T. cruzi has been a major problem with treating Chagas disease and similar resistance could evolve to molecules that are delivered by paratransgenic mechanisms. Use of multiple effector molecules would help reduce development of resistance. AMPs target cellular membranes and internal components to exert toxicity while β-glucanase works on sugar moieties sequestered on the surface of the parasite. Dual targeting of T. cruzi should not only slow resistance but also permit synergistic or additive effects on T. cruzi lethality. We have also developed recombinant single chain antibodies that further strengthen the tool chest . These single chain antibodies, which target sialic acid epitopes on T. cruzi surfaces, will be used in an integrated and comprehensive field-based strategy to target vector stages of T. cruzi, as part of paratransgenic control of Chagas disease.
Currently, we are conducting in vivo co-infection studies in R. prolixus nymphs using engineered lines of R. rhodnii to study the effects of our molecules on the arthropods and verify efficacy. Aposymbiotic nymphs are infected with transformed R. rhodnii, allowed to develop, and challenged with T. cruzi in a blood meal. We have set up in vivo studies that examine the effects of AMP combinations and recombinant β-glucanase on arthropod maturation, fecundity and competence to carry and permit maturation of T. cruzi. Successful colonization of R. prolixus nymphs by the transgenic symbionts and elimination of T. cruzi carriage in all paratransgenic insects will set the stage for simulated field trials.
Chagas disease remains a public health concern and an important neglected disease. Despite efforts by governments and overall improvement in living conditions, detection, and treatments, tens of millions of people are exposed to the disease-causing parasites annually. Current methods involving insecticides as a method of vector control work only in small domestic areas and carry high costs. The paratransgenic method using recombinant symbiotic bacteria may serve as a valuable adjunct in the ongoing battle to eliminate Chagas disease.
Tris-buffered saline with 0.5% tween-20.
This work was supported by NIH/NIAID RO1AI66045 (RD). CJ was support by an ARRA Administrative Supplement for Summer Research Experience for Students, grant number 3RO1AI066045-04S1.
CJ, NK, SW, AF and IH contributed to all the experiments described in this paper. CJ wrote the manuscript. AF, IH and RD analyzed and interpreted the results. IH and RD finalized the manuscript in consultation with the other authors. All authors revised and approved the final version of this manuscript.


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Voter questionnaire
As the Culture Cloud project draws to its conclusion, the evaluation team are eager to gain as much feedback about the experience as possible from participants.
On one hand this entails questioning as many of the 900+ competition entrants as possible and to this end we've mailed all of them with a questionnaire and invitation to feedback to us in detail.
On the other hand we are interested in finding out as much about as many of the voters as possible. There are over 40000 registered 'likes' for the shortlisted artworks posted on Artfinder which indicates that the project has attracted a lot of attention. We've launched an online questionnaire for voters and encourage each and every one of them to take a minute or two to answer the few questions we've set.
The voter questionnaire can be found here.
Respondents are welcome to share this link and to feedback to us directly – via this site, or to paul.long@bcu.ac.uk.
Thanks for your participation!
Thinking about online art and the space of the gallery
Since encountering Culture Cloud and the nature of originating partner organizations New Art Exchange and Artfinder, a core question for the evaluation team concerns the very nature of art online, its definition, mode of consumption and, indeed, the expectations of all involved in the relationships forged around artworks.
Here, we can identify a number of ways of thinking about art and the online world related to this project that will be helpful in evaluating its R&D dimensions and the nature of any innovation in the relationship between gallery, art, artist and consumer.
Online Cultural Democracy
One aspect of the approach that we have taken to evaluating the Culture Cloud is to place it in the context of other online projects that have evinced similar aims, objectives and methods. One such project is Democracy which came to my attention as a result of a presentation by Jim Richardson, founder and director of Sumo Design and MuseumNext, Europe's major conference on social media and museums.
Richardson's presentation took place at one of the AHRC-funded Digital Transformations Workshops organized by the University of Westminster: 'a research network exploring digital transformations in the creative relationships between cultural and media organisations and their users.' Clearly, an initiative that is a potentially important point of reference for Culture Cloud and related discussions.
The Culture Cloud Goes Live!
The call for artists to upload their work for consideration as part of the Culture Cloud project is now online at: theculturecloud.com or theculturecloud.co.uk. There is also a Facebook page dedicated to circulating information on the project. The Twitter feed can be found at: @NaeCultureCloud.
Potential contributors were invited to start uploading/registering art works from 5 March 2012. Registration is open until midnight Friday 4 May 2012.
Partner Profile: What is (who are) Artfinder?
Artfinder describes a technology, 'an exciting new concept for discovering, experiencing and sharing art' which 'gives you access to hundreds of thousands of paintings', allowing you to build 'your own art profile'. It promises to be the Last.fm of art, offers to sell you prints of most of the artwork shown, and is most emphatically not a museum.
The Evaluation Team
The academic evaluation team on the Culture Cloud project comprises Dr Nick Webber, Dr Simon Barber, Dr. Paul Long and Professor Tim Wall. They are representatives of the Interactive Cultures research team, based in the Birmingham Centre for Media & Cultural Research, which focuses on work associated with the cultural and creative industries, emphasising cultural consumption and new digital technologies.
The team have run externally funded projects and published work which deepens and integrates the following themes:
cultural practice, history and heritage;
social media, interactivity, digital participation;
digital and online archives;
locative-media, gaming cultural practices, application to other domains.
Partner Profile: What is 'The New Art Exchange'?
New Art Exchange is a visual arts organisation based in the Hyson Green area of Nottingham, housed in a RIBA-award winning building designed by the architects Hawkins Brown.
Formed in September 2003, NAE was conceived as an organisation that would steer and manage the development of Nottingham's first dedicated cultural facility for Black contemporary arts. This conception can be understood as result of NAE's formation out of a partnership between APNA Arts and EMACA Visual Arts. APNA Arts (Apna is Hindi for 'ours') focused on South Asian arts and played a key role in the development of the 'Nottingham Mela' in 2008. EMACA – East Midlands African Caribbean Arts – was an independent organisation that existed to promote the work of local artists of African and African- Caribbean heritage within the East Midlands employing various media through the use of workshops, exhibitions, performances and discussions.
The project: What is 'Culture Cloud'?
The 'Culture Cloud' project was successfully pitched to NESTA/ACE/AHRC in response to the 'Digital Research and Development' funding call. 'Culture Cloud' is a partnership between the Nottingham-based visual arts organisation New Art Exchange and Artfinder acting as technology provider. The team from Birmingham Centre for Media and Cultural Research will act as academic evaluators on this project.
Artfinder is an ambitious internet start-up that brings the approach of social media practice to the world of art. Describing Artfinder, the Guardian's Jemima Kiss suggests that we think 'of movie details site IMDB, music recommendations service Last.fm, gig tracking site Songkick, online mixture site Muxtape (RIP) and Shazam, the music identification service all as shorthand for some aspect of ArtFinder is trying to do. (See: Jemima Kiss, 'ArtFinder brings social media to art', 6 May 2011, http://www.guardian.co.uk/media/pda/2011/may/06/artfinder-social-media-art [accessed 3/2/12]).


Big data analytics company Teradata has unveiled a next generation analytics solution which will enable data-driven health care.
According to Teradata, the framework enables health care payers and providers to "harness new analytical capabilities to reduce costs, improve customer engagement and incentives, better coordinate quality care and support risk-finance transparency".
Based on new capabilities in the Teradata unified data architecture, the Teradata healthcare data model, and Teradata's healthcare applications from the Aster Applications Center, the framework includes analytics for paths to surgery, admission diagnosis procedure paths, patient affinity, patient compare, impact analysis, and drug prescription affinity analysis.
"More timely analytic insight and customer communications have powerful benefits that enhance multiple dimensions of intelligence around patient care," said Scott Sobers, director, healthcare and life sciences industry marketing, Teradata.
"For instance, a healthcare organization that seeks to conduct proactive intervention to help prevent re-admissions can be a challenge. Today, the process takes too long; it's inefficient and costly. With the healthcare framework in place, organizations can reduce unnecessary re-admissions - better serving patients and ensuring a more economical process for the business," he added.
Judy Hanover, research director at IDC Health Insights, said: "The healthcare sector is investing significantly in BI and analytics, as the technologies are important enablers of success in accountable care delivery.
"Most providers have had mixed results from past investments, and are intensifying their efforts to select strong tools that deliver actionable results from analytics. Analytics can help hospitals and health systems improve visibility into risk, financials, and customer engagement."


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Mount Calvary is een plaats (village) in de Amerikaanse staat Wisconsin, en valt bestuurlijk gezien onder Fond du Lac County.

Demografie
Bij de volkstelling in 2000 werd het aantal inwoners vastgesteld op 956. In 2006 is het aantal inwoners door het United States Census Bureau geschat op 939, een daling van 17 (-1,8%).

Geografie
Volgens het United States Census Bureau beslaat de plaats een oppervlakte van 2,7 km², geheel bestaande uit land.

Plaatsen in de nabije omgeving
De onderstaande figuur toont nabijgelegen plaatsen in een straal van 20 km rond Mount Calvary.

Externe link

Plaats in Wisconsin

redpajama_set_name: RedPajamaWikipedia


text:
Q: Implicit function equation $\frac{\partial z}{\partial x}$ calculation $ xz^2+e^{yz}=0 $
I know this equation:
$ 0=Fx \frac{\partial x}{\partial x} + Fy \frac{\partial y}{\partial x} + Fz \frac{\partial z}{\partial x}$
now I transform it to:
$\frac{\partial z}{\partial x}=\frac{-Fx-Fy\frac{\partial y}{\partial x}}{Fz} $
and I get equation
$\frac{\partial z}{\partial x}=\frac{-Fx}{Fz}=\frac{z^2}{2xz+e^{yz}y} $
Can someone explain why $ Fy\frac{\partial y}{\partial x}=0$ ?
I think when we are taking $\frac{\partial z}{\partial x} $ we are treating y as a constant so Fy=0?

A: Partial derivative notation is ambiguous. But you've correctly identified the crucial missing piece of information that was intended in your calculation — all of the derivatives are meant to be computed subject to the restriction that $y$ is constant.
It's not $F_y$ that's zero, however; $F_y$ still equals $ze^{yz}$. It's $\frac{\partial y}{\partial x}$ that's zero — it's zero because the intent of the calculation is to take derivatives while holding $y$ constant.
Note that if the intent was to take derivatives while holding $z$ constant, then $\frac{\partial y}{\partial x}$ would not be zero.
One notation I've seen for adding in the missing information is
$$0=F_x \left.\frac{\partial x}{\partial x}\right|_{y \text{ const}} + F_y \left.\frac{\partial y}{\partial x}\right|_{y \text{ const}}  + F_z \left.\frac{\partial z}{\partial x}\right|_{y \text{ const}} $$
or some abbreviation thereof.

For the sake of precision, I should point out what $F_y$ means too. If we write down the function $F(a,b,c) = a c^2 + e^{bc}$  (so that the given equation is the value of $F(x,y,z)$), $F_y$ is shorthand for $F_2(x, y, z)$, where $F_2$ means to take the derivative of $F_2$ in its second place.


To talk about an essential component of anything, we can use metaphors involving transportation or architecture: a linchpin keeps the wheel on the axle or, figuratively, keeps everything rolling along, much like a k_______ keeps all the other stones in place or, figuratively, keeps everything together.
A literal linchpin is a pin or peg that keeps a wheel attached to its axle.
A figurative linchpin is a very important part of something.
None, but some people use it loosely as an adjective: "the linchpin policy," "this linchpin issue."
A linchpin is sometimes a person: "She's the linchpin in her group of friends."
Usually, though, a linchpin is a thing: "Merchandise is the linchpin of the filmmaker's financial success."
More specifically, a linchpin is often an abstract thing, like a concept, a quality, a process, or a principle: "kindness is the linchpin of his argument," "one-on-one lessons are the linchpin of our success," "tolerating differences and diversity is the linchpin of our democracy."
You usually call something the linchpin of something else, but you can also have a linchpin in a situation or a linchpin for someone or something.
The linchpin of his argument is our need to reduce the national debt, but for many listeners that goal is abstract, even negligible.
Accurate observation isn't a footnote in the scientific method; it's a linchpin.
Look away from the screen to explain the definition in your own words. You'll know you understand what "linchpin" means when you can explain it without saying "essential component" or "vital thing."
What's the key element in your next big dream, goal, project, or plan? Fill in the blank: "I need to get the linchpin in place: _____."
Example: "We need to get the linchpin in place: the discipline to start cooking a meal before we're already hungry."
Try that again today with the letters BLRTAEEO. Draw three vertical and three horizontal lines on some scrap paper, and go for it!
2. By cancelling the traditional Nutcracker shows, they've _____ the linchpin of the winter season.


When we examined the likely outcomes of next week's FOMC meeting yesterday [see: Now and Then] we devoted a couple of column inches to the possibility that Wall Street would throw a fit, much like it did in October 2014, January 2015 and August 2015, to demonstrate how sensitive it was to the taking away of the proverbial punch bowl.
Having said all that, there is one other possibility that shouldn't be completely ignored. As we've seen in the past, a sudden, severe plunge in stocks can be very effective in changing the FOMC's plans.
Were SPX to drop to, say, 1920, 1856 or 1823 in the next several sessions (6.4, 9.6 or 11.2%), there's a pretty good chance that the Fed would delay actual implementation of a rate hike.
The words seemed rather silly as I typed them, particularly since every tiny dip was immediately bought yesterday. With another of those now-routine 1% pre-opening plunges facing us this morning, they seem a little less silly.


Aluminium montageplaatset met b-kogel afmeting 15,9 x 5,1 cm, voorzien van diverse montagegaten. RAM Mounts B-kogel rond en 9 cm klemhouder maken de set compleet voor directe montage.
The RAM-B-111 consists of a double socket arm, 2.5" diameter round base and 6.25" x 2" rectangle base. The round base contains pre-drilled holes, including the universal AMPS hole pattern. The rectangle base contains different combinations of pre-drilled holes which will match up to many electronic devices and mounts, including Marine GPS, CB Radios, Ham Radios, Gimbal Brackets and more.


The Bluebeard tale reset in a 19th century cult waiting for the end of days in upstate NY. On the plus side, this is good and creepy. The author captures the atmosphere of upstate NY really well. In fact I will admit that may be that he captured it well enough to make me uncomfortable and dislike parts of the book just because I live in NY state and I don't want to be afraid to go into the woods.
The cult is well portrayed too - the evil charismatic preacher in his utter self assurance is one scary guy. His followers may be even scarier, so defeated that they are actually glad to hear that the world is ending and they don't have to keep trying to make sense of it, so eager to turn over their decisionmaking to a charismatic who looks like he might have some answers.
On the minus side, the story is a tale of three sisters - but a lot of time is spent with the first sister, less with the second, and even less with the third. The balance didn't work so well for me. I could have done with a little less time listening to Vern go over the same ground repeatedly, and a little more getting to know Kate.
The major difficulty for me was that somehow the balance between the natural and supernatural elements didn't quite work. Gothic horror is hard to write, it is supposed to be inchoate, you're supposed to spend a lot of the story wondering if the central character is seeing ghosts or just going insane. Its not easy to make that work, and here it doesn't always.
A lot of the book was so extremely realistic in its tone, very specific descriptions of architecture and clothing and place names, and landscape. Because of that level of realism I felt like I wanted the gothic horror elements to be more grounded too. For example, its summertime - I don't understand how the chamber of blood doesn't clot, or smell, or draw flies. I don't understand what the ghosts are doing the third floor or what their role is in the story. I don't know who the Angel of Darkness is, or even if it exists. I'm left with too many questions.
Its still a good, scary, well written story, it just feels like it came really close to being something even more interesting but in the end it didn't quite gell.