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ry.easing[this.options.easing||(jQuery.easing.swing?"swing":"linear")](th=
is.state,n,0,1,this.options.duration);this.now=3Dthis.start+((this.end-th=
is.start)*this.pos);this.update();}return =
true;}};jQuery.extend(jQuery.fx,{speeds:{slow:600,fast:200,def:400},step:=
{scrollLeft:function(fx){fx.elem.scrollLeft=3Dfx.now;},scrollTop:function=
(fx){fx.elem.scrollTop=3Dfx.now;},opacity:function(fx){jQuery.attr(fx.ele=
m.style,"opacity",fx.now);},_default:function(fx){fx.elem.style[fx.prop]=3D=
fx.now+fx.unit;}}});jQuery.fn.offset=3Dfunction(){var =
left=3D0,top=3D0,elem=3Dthis[0],results;if(elem)with(jQuery.browser){var =
parent=3Delem.parentNode,offsetChild=3Delem,offsetParent=3Delem.offsetPar=
ent,doc=3Delem.ownerDocument,safari2=3Dsafari&&parseInt(version)<522&&!/a=
dobeair/i.test(userAgent),css=3DjQuery.curCSS,fixed=3Dcss(elem,"position"=
)=3D=3D"fixed";if(elem.getBoundingClientRect){var =
box=3Delem.getBoundingClientRect();add(box.left+Math.max(doc.documentElem=
ent.scrollLeft,doc.body.scrollLeft),box.top+Math.max(doc.documentElement.=
scrollTop,doc.body.scrollTop));add(-doc.documentElement.clientLeft,-doc.d=
ocumentElement.clientTop);}else{add(elem.offsetLeft,elem.offsetTop);while=
(offsetParent){add(offsetParent.offsetLeft,offsetParent.offsetTop);if(moz=
illa&&!/^t(able|d|h)$/i.test(offsetParent.tagName)||safari&&!safari2)bord=
er(offsetParent);if(!fixed&&css(offsetParent,"position")=3D=3D"fixed")fix=
ed=3Dtrue;offsetChild=3D/^body$/i.test(offsetParent.tagName)?offsetChild:=
offsetParent;offsetParent=3DoffsetParent.offsetParent;}while(parent&&pare=
nt.tagName&&!/^body|html$/i.test(parent.tagName)){if(!/^inline|table.*$/i=
.test(css(parent,"display")))add(-parent.scrollLeft,-parent.scrollTop);if=
(mozilla&&css(parent,"overflow")!=3D"visible")border(parent);parent=3Dpar=
ent.parentNode;}if((safari2&&(fixed||css(offsetChild,"position")=3D=3D"ab=
solute"))||(mozilla&&css(offsetChild,"position")!=3D"absolute"))add(-doc.=
body.offsetLeft,-doc.body.offsetTop);if(fixed)add(Math.max(doc.documentEl=
ement.scrollLeft,doc.body.scrollLeft),Math.max(doc.documentElement.scroll=
Top,doc.body.scrollTop));}results=3D{top:top,left:left};}function =
border(elem){add(jQuery.curCSS(elem,"borderLeftWidth",true),jQuery.curCSS=
(elem,"borderTopWidth",true));}function =
add(l,t){left+=3DparseInt(l,10)||0;top+=3DparseInt(t,10)||0;}return =
results;};jQuery.fn.extend({position:function(){var =
left=3D0,top=3D0,results;if(this[0]){var =
offsetParent=3Dthis.offsetParent(),offset=3Dthis.offset(),parentOffset=3D=
/^body|html$/i.test(offsetParent[0].tagName)?{top:0,left:0}:offsetParent.=
offset();offset.top-=3Dnum(this,'marginTop');offset.left-=3Dnum(this,'mar=
ginLeft');parentOffset.top+=3Dnum(offsetParent,'borderTopWidth');parentOf=
fset.left+=3Dnum(offsetParent,'borderLeftWidth');results=3D{top:offset.to=
p-parentOffset.top,left:offset.left-parentOffset.left};}return =
results;},offsetParent:function(){var =
offsetParent=3Dthis[0].offsetParent;while(offsetParent&&(!/^body|html$/i.=
test(offsetParent.tagName)&&jQuery.css(offsetParent,'position')=3D=3D'sta=
tic'))offsetParent=3DoffsetParent.offsetParent;return =
jQuery(offsetParent);}});jQuery.each(['Left','Top'],function(i,name){var =
method=3D'scroll'+name;jQuery.fn[method]=3Dfunction(val){if(!this[0])retu=
rn;return =
val!=3Dundefined?this.each(function(){this=3D=3Dwindow||this=3D=3Ddocumen=
t?window.scrollTo(!i?val:jQuery(window).scrollLeft(),i?val:jQuery(window)=
.scrollTop()):this[method]=3Dval;}):this[0]=3D=3Dwindow||this[0]=3D=3Ddoc=
ument?self[i?'pageYOffset':'pageXOffset']||jQuery.boxModel&&document.docu=
mentElement[method]||document.body[method]:this[0][method];};});jQuery.ea=
ch(["Height","Width"],function(i,name){var =
tl=3Di?"Left":"Top",br=3Di?"Right":"Bottom";jQuery.fn["inner"+name]=3Dfun=
ction(){return =
this[name.toLowerCase()]()+num(this,"padding"+tl)+num(this,"padding"+br);=
};jQuery.fn["outer"+name]=3Dfunction(margin){return =
this["inner"+name]()+num(this,"border"+tl+"Width")+num(this,"border"+br+"=
Width")+(margin?num(this,"margin"+tl)+num(this,"margin"+br):0);};});})();
------=_NextPart_000_0000_01C99F0E.BB8D06B0
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www3.interscience.wiley.com/freeflow/js/genlink.js

function get_link(a, b) {=0A=
	document.write("<a href=3D\"/cgi-bin" + a + "\" target=3D\"_top\">" + b =
+ "</a>");=0A=
}=0A=
=0A=
function get_pdf_link(a, b, c) {=0A=
	document.writeln('<a href=3D"/cgi-bin' + a + '" target=3D"_top">' + b + =
'</a> (Size: ' + c + 'K)');=0A=
}=0A=
=0A=
function get_button_link(cgi, params, imgbutton) {=0A=
	document.writeln('<a href=3D"/cgi-bin/' + cgi + params + '" =
target=3D\"_top\"><img src=3D"' + imgbutton + '" border=3D0 alt=3D"Click =
to Access Graphical Table of Contents"></a>');=0A=
}=0A=
=0A=
function get_text_link(url, linktext)=0A=
{=0A=
	document.writeln('<a href=3D"' + url + '" target=3D\"_top\">' + =
linktext + '</a>');=0A=
}=0A=
=0A=
function get_htmlftext_link(a, b) {=0A=
	document.writeln("<a href=3D\"/cgi-bin" + a + "\" target=3D\"_top\">" + =
b + "</a>, ");=0A=
}=0A=
=0A=

------=_NextPart_000_0000_01C99F0E.BB8D06B0
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www3.interscience.wiley.com/freeflow/js/personalisation.js

/*=0A=
* Project				:	Interscience Redesign=0A=
* File					:	personalisation.js=0A=
* Creation Date			:	25th March 2003=0A=
* Author				:	Jonathan Sugar=0A=
*=0A=
* Various functions to deliver personalisation functionality.=0A=
*/=0A=
=0A=
// Pre-declare userName so it's available when we need to test if the =
page=0A=
// assembler ran OK=0A=
var userName =3D null;=0A=
var shcrtCookie =3D null;=0A=
=0A=
// Browser detect globals:=0A=
var uaStr =3D navigator.userAgent.toLowerCase();=0A=
var uaVer =3D parseInt(navigator.appVersion);=0A=
// for detecting netscape 4=0A=
var netscape =3D (uaStr.indexOf("mozilla") > -1);=0A=
var netscape4 =3D (netscape && uaVer < 5);=0A=
=0A=
// Factory Methods=0A=
=0A=
// Simple wraper object to encapsulate the data for a co-branding item.=0A=
function coBrandingItem(image, imageAltText, href)=0A=
{=0A=
	//alert("Making new coBrandingItem:\nImage: " + image + =
"\nimageAltText" + imageAltText + "\nhref: " + href);=0A=
	this.image =3D image;=0A=
	this.imageAltText =3D imageAltText;=0A=
	this.href =3D href;=0A=
}=0A=
=0A=
/*=0A=
* Insert the 'Customer co-branding' section.=0A=
* -----------------------------=0A=
*=0A=
* Author: Jonathan Sugar, March 2003.=0A=
*=0A=
* 1.	Insert customer co branding. Loop through 'customerCoBranding' array=0A=
* 		inserting XHTML where needed.=0A=
*=0A=
* NOTE: the array contains ONLY the resource location (the location of a =
gif=0A=
* on the file system), the required XHTML is then added where needed. =
This way=0A=
* we maintain the seperation between presentation and content.=0A=
*/=0A=
function insertCustomerCoBranding(maxBranding)=0A=
{=0A=
	if( customerCoBranding[0].image !=3D "none")=0A=
	{=0A=
		document.writeln('<div id=3D"providedBy" class=3D"item">');=0A=
		document.writeln('<h5><img src=3D"/images/txt.provided-by.gif" =
width=3D"71" height=3D"16" alt=3D"provided by" border=3D"0" /></h5>');=0A=
		document.writeln('<ul class=3D"noBullet">');=0A=
=0A=
		var i;=0A=
		//Change start for bug#2288=0A=
		if(typeof(maxBranding) =3D=3D "undefined" | maxBranding =3D=3D "" | =
maxBranding =3D=3D 0){=0A=
			maxBranding =3D 3; // DEFAULT Value for No of Cobranding dispaly=0A=
		}=0A=
		var size =3D customerCoBranding.length;=0A=
		if(maxBranding !=3D -1 & size-1 > maxBranding){=0A=
			size =3D maxBranding;=0A=
		}=0A=
		//Change End for bug#2288=0A=
=0A=
		// Add the co-branding images o the page.=0A=
		//for(i=3D0; i < customerCoBranding.length; i++ )=0A=
		for(i=3D0; i < size; i++ )=0A=
		{=0A=
			// ignore the terminating array element.=0A=
			if(customerCoBranding[i].image =3D=3D "none")=0A=
			{=0A=
				continue;=0A=
			}=0A=
=0A=
			this.image =3D customerCoBranding[i].image;=0A=
			this.imageAltText =3D customerCoBranding[i].imageAltText;=0A=
			this.href =3D customerCoBranding[i].href;=0A=
=0A=
			if (image =3D=3D "")=0A=
			{=0A=
				if(href =3D=3D "")=0A=
				{=0A=
					document.writeln('<li>' + imageAltText + '</li>');=0A=
				}=0A=
				else=0A=
				{=0A=
					document.writeln('<li><a href=3D"' + href + '">' + imageAltText + =
'</a></li>');=0A=
				}=0A=
			}=0A=
			else=0A=
			{=0A=
				if(href =3D=3D "")=0A=
				{=0A=
					document.writeln('<li><img src=3D"' + image + '" alt=3D"' + =
imageAltText + '" border=3D"0" /></li>');=0A=
				}=0A=
				else=0A=
				{=0A=
					document.writeln('<li><a href=3D"' + href + '"><img src=3D"' + =
image + '" alt=3D"' + imageAltText + '" border=3D"0" /></a></li>');=0A=
				}=0A=
			}=0A=
		}=0A=
=0A=
		// If there are more co-branding images add in a link to them.=0A=
		// Changes made for Bug#2288=0A=
		//if(moreCustomerCoBranding =3D=3D "true")=0A=
		if(maxBranding !=3D -1 & customerCoBranding.length-1 > maxBranding)=0A=
		{=0A=
			document.writeln('<li><a href=3D"/cgi-bin/myprofile" =
class=3D"linkMore">More Providers</a></li>');=0A=
		}=0A=
=0A=
		document.writeln('</ul>');=0A=
		document.writeln('</div>');=0A=
		document.writeln('<div class=3D"line"><img src=3D"/images/dot.CCC.gif" =
width=3D"100%" height=3D"1" alt=3D"" border=3D"0" /></div>');=0A=
	}=0A=
}=0A=
=0A=
/*=0A=
* Insert the 'My Area' section.=0A=
* -----------------------------=0A=
*=0A=
* Author: Jonathan Sugar, March 2003.=0A=
* Updated: Ben Marvell, April 2006=0A=
*=0A=
* My profile link absolute as this file isnt only run from w3 anymore.=0A=
*/=0A=
function insertMyArea(userdata)=0A=
{=0A=
	// just in case the page assembler isn't running.=0A=
	=0A=
	=0A=
	if (userName !=3D null)=0A=
	{=0A=
		document.writeln("<ul>");=0A=
		if(typeof(environment)!=3D'undefined' && environment!=3D'PRODUCTION')=0A=
		{=0A=
			if(userName !=3D "")=0A=
			{=0A=
				//write out the user specific data for the menu=0A=
				if (userdata !=3D null){=0A=
					if (userName.length > 33){=0A=
						var writedata =3D "<li id=3D\"user\"><strong>" + =
userName.substring(0,33) + "...</strong>, access your</li>";=0A=
					} else {=0A=
						var writedata =3D "<li id=3D\"user\"><strong>" + userName + =
"</strong>, access your</li>";=0A=
					}=0A=
	=0A=
					for (var x=3D0; x<userdata.length; x++)=0A=
					{=0A=
						writedata +=3D userdata[x];=0A=
					}=0A=
					document.writeln(writedata);=0A=
				}=0A=
				document.writeln(getShCrtLink(true));=0A=
			}=0A=
			else =0A=
				document.writeln(getShCrtLink(false));=0A=
				=0A=
			toggleShCrtLink();=0A=
		=0A=
			document.writeln("<li><a href=3D'/cgi-bin/myprofile' target=3D'_top' =
class=3D'menuSideArrow'>My Profile</a></li>");	=0A=
		}		=0A=
		document.writeln(getLoginOutLink());=0A=
		//CR 3652 getAthenStatus() as added=0A=
		document.writeln(getAthenStatus());=0A=
		document.writeln("</ul>");=0A=
	} else {=0A=
		if(typeof(environment)!=3D'undefined' && =
environment=3D=3D'PRODUCTION') {=0A=
			document.writeln("<ul>");=0A=
			if (userdata !=3D null){=0A=
				var writedata =3D "";=0A=
				for (var x=3D0; x<userdata.length; x++) {=0A=
					writedata +=3D userdata[x];=0A=
				}=0A=
				document.writeln(writedata);=0A=
			}=0A=
			document.writeln("<li><a href=3D'/prodinterface/sulogout' =
target=3D'_top' class=3D'menuSideArrow'>Log Out</a></li>");=0A=
			document.writeln("</ul>");	=0A=
		}=0A=
	}=0A=
	if (menuloadedflag){=0A=
		wisMenu();=0A=
	}=0A=
}=0A=
=0A=
/*=0A=
* Insert the Customer Admin 'Login/Logout' link=0A=
* -----------------------------=0A=
*=0A=
* Author: Bobby Jack, December 2004.=0A=
*=0A=
*/=0A=
function insertAdminLink(page)=0A=
{=0A=
	val =3D getCookie("AID");=0A=
	=0A=
	if (val && val !=3D "-1")=0A=
	{=0A=
		document.writeln('<li class=3D"menuSideArrow" style=3D"padding-left: =
80px;"><a class=3D"menuSideArrow" =
href=3D"/cgi-bin/logoutcustadmin?Page=3D/cgi-bin/bookpicker">Logout</a></=
li>');=0A=
	}=0A=
	else=0A=
	{=0A=
		document.writeln('<li class=3D"menuSideArrow" style=3D"padding-left: =
80px;"><a class=3D"menuSideArrow" =
href=3D"/cgi-bin/custnavlogin?TPL=3Dcustomer-admin.customer&Page=3D' + =
page + '">Login</a></li>');=0A=
	}=0A=
}=0A=
=0A=
/*=0A=
* Insert the username section.=0A=
* -----------------------------=0A=
*=0A=
* Author: Jonathan Sugar, March 2003.=0A=
*=0A=
* 1. Insert the username.=0A=
*/=0A=
function insertUserName()=0A=
{=0A=
	document.write(userName);=0A=
}=0A=
=0A=
/*=0A=
* Return the appropriate link to the login/logout cgi=0A=
* ---------------------------------------------------=0A=
*=0A=
* Author: Jonathan Sugar, March 2003.=0A=
* Updated: Ben Marvell, April 2006=0A=
* Had to change the links absolute as this file isnt only run from w3 =
anymore.=0A=
*/=0A=
function getLoginOutLink()=0A=
{=0A=
	if(userName !=3D "")=0A=
	{=0A=
		return "<li><a href=3D'/cgi-bin/logout' target=3D'_top' =
class=3D'menuSideArrow'>Log Out</a></li>";=0A=
	}else=0A=
	{=0A=
		return "<li><a href=3D'/' target=3D'_top' class=3D'menuSideArrow'>Log =
In</a></li>";=0A=
	}=0A=
}=0A=
=0A=
/*=0A=
* Return the appropriate link to the shopping cart content page=0A=
* ---------------------------------------------------=0A=
*=0A=
* Author: Zhiming & Warrell June 2007=0A=
*/=0A=
function getShCrtLink(logged)=0A=
{=0A=
	if(logged =3D=3D true )=0A=
		return "<li><div ID=3D'shcrt_id'><a href=3D'/cart/shoppingcart' =
target=3D'_top' class=3D'menuSideArrow'><img src=3D'/images/cart.gif' =
border=3D'0' align=3D'absbottom'/>My Cart</a></div></li>";=0A=
	else=0A=
		return "<li id=3D'mycart'><div ID=3D'shcrt_id'><a =
href=3D'/cart/shoppingcart' target=3D'_top' class=3D'menuSideArrow'><img =
src=3D'/images/cart.gif' border=3D'0' align=3D'absbottom'/>My =
Cart</a></div></li>";=0A=
}=0A=
=0A=
function toggleShCrtLink()=0A=
{=0A=
	shcrtCookie =3D readCookie("SHCRT");=0A=
	if(shcrtCookie !=3D null)=0A=
	{=0A=
		if(document.all)=0A=
		{=0A=
			return true;=0A=
		}=0A=
		else=0A=
		{=0A=
			document.getElementById("shcrt_id").style.display =3D "block";=0A=
		}=0A=
	}=0A=
	else=0A=
	{=0A=
		if(document.all)=0A=
		{=0A=
			return true;=0A=
		}=0A=
		else=0A=
		{=0A=
			document.getElementById("shcrt_id").style.display =3D "none";=0A=
		}=0A=
	}=0A=
}=0A=
=0A=
=0A=
=0A=
function readCookie(name)=0A=
{=0A=
	var nameEQ =3D name + "=3D";=0A=
	var ca =3D document.cookie.split(';');=0A=
	for(var i=3D0; i<ca.length;i++)=0A=
	{=0A=
		var c =3D ca[i];=0A=
		while (c.charAt(0)=3D=3D' ') c =3D c.substring(1,c.length);=0A=
			if (c.indexOf(nameEQ) =3D=3D 0) return =
c.substring(nameEQ.length,c.length);=0A=
	}=0A=
	return null;=0A=
}=0A=
=0A=
/*=0A=
* Author: Aung Kyaw Win, March 2008.=0A=
* CR 3652: To check Athen Login Status =0A=
* Bug 5262 To redirect to speicifc publication page=0A=
*/=0A=
function getAthenStatus()=0A=
{=0A=
	if (readCookie('Augment')=3D=3D"Athens")=0A=
	{=0A=
	 	return '<li class=3D"menuDOM" id=3D"user"><strong>You are logged in =
to Athens</strong></li>';=0A=
	}=0A=
	else=0A=
	{=0A=
		return "<li><a =
href=3D'/cgi-bin/athenslogin?Page=3D"+location.pathname+location.search+"=
' target=3D'_top' class=3D'menuSideArrow'>Athens Log In</a></li>";=0A=
	}=0A=
	=0A=
}
------=_NextPart_000_0000_01C99F0E.BB8D06B0
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www3.interscience.wiley.com/freeflow/js/site.js

/************************************************************************=
***************************=0A=
 	FILE:		site.js=0A=
 	AUTHOR: 	Jonathan Sugar, Chris Neatby-Smith and Digital Pulp. July =
2003.=0A=
 =0A=
 	DESCRIPTION:	Javascript relating to site wide functionality across =0A=
 					InterScience.=0A=
 =0A=
 	INCLUDES FUNCTIONS=0A=
 	=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=0A=
 	=0A=
 	Search Functions=0A=
 	----------------=0A=
 	=0A=
 	selectAndSubmit() - for deciding where to submit a query based on the =
state of the form.=0A=
 	processallContentSearch() - for validating form input for an all =
content search.=0A=
 	processTitleSearch() - for validating form input for a Title search.=0A=
 	checkCitationSearch() - for validating the citation search form.=0A=
 	selectAll() - check all the checkboxes in a form.=0A=
	do_popup() - stasa Acronym Finder window popup function.=0A=
	=0A=
 	=0A=
 	Other Functions=0A=
 	---------------=0A=
 	=0A=
 	fixInlineLists() - 	fix netscape 4.x bug where <li> tags with images =
display both an image and=0A=
 						the default bullet. 	=0A=
=0A=
 =
*************************************************************************=
**************************/=0A=
=0A=
var http =3D false;=0A=
if(navigator.appName =3D=3D "Microsoft Internet Explorer") {=0A=
  http =3D new ActiveXObject("Microsoft.XMLHTTP");=0A=
} else {=0A=
  http =3D new XMLHttpRequest();=0A=
} =0A=
=0A=
// Global Variables=0A=
var debug =3D false;=0A=
var first =3D true;=0A=
=0A=
// Browser detect globals:=0A=
var uaStr =3D navigator.userAgent.toLowerCase();=0A=
var uaVer =3D parseInt(navigator.appVersion);=0A=
// for detecting netscape 4=0A=
var netscape =3D (uaStr.indexOf("mozilla") > -1);=0A=
var netscape4 =3D (netscape && uaVer < 5);=0A=
=0A=
/*=0A=
 * Used to switch search mode between APS and TitleFinder on quicksearch =
boxes=0A=
 * =
-------------------------------------------------------------------------=
--=0A=
 *=0A=
 * 1.	Decides which search is required based on the value of the radio =
buttons=0A=
 *		in 'document.formSelector.whichForm'.=0A=
 *=0A=
 * 2.	Calls individual functions to process the forms and submit them.=0A=
 *=0A=
 */=0A=
function selectAndSubmit()=0A=
{=0A=
	var formSelection;=0A=
=0A=
	// find out which form we need to process.=0A=
	var selector =3D document.formSelector.whichForm;=0A=
	for (var i =3D 0; i < selector.length; i++){=0A=
		if (selector[i].checked){=0A=
			formSelection =3D selector[i].value;=0A=
		}=0A=
	}=0A=
	=0A=
	// Call the relevent function to process the required form.=0A=
	if(formSelection =3D=3D "allContentSearch"){=0A=
		processallContentSearch(document.allContentSearchForm, =
document.formSelector)=0A=
	}=0A=
	else if(formSelection =3D=3D "searchTitles"){=0A=
		processTitleSearch(document.searchTitles, document.formSelector)=0A=
	}=0A=
	else{=0A=
		return false;=0A=
	}=0A=
	=0A=
	//@@@DM It was anoying me this error		=0A=
	return false;=0A=
}=0A=
=0A=
/*=0A=
*Process the ADS search form=0A=
*=0A=
*/=0A=
function trim(str)=0A=
{=0A=
    return str.replace(/(^\s*)|(\s*$)/g,"");=0A=
}=0A=
=0A=
function ADSquery(t) {=0A=
    var keyword =3D trim(t.getElementsByTagName("input")[0].value);=0A=
    if (keyword!=3D"") {=0A=
	    =
window.open("","popup","toolbar=3Dyes,location=3Dyes,status=3Dyes,menubar=
=3Dno,scrollbars=3Dyes,resizable=3Dyes,width=3D800,height=3D500");=0A=
		t.target=3D"popup";=0A=
		t.qsearch.value=3Dkeyword;=0A=
		return true;=0A=
		=
//processADSSearch(document.ADSSearchForm,document.ADSQueryForm,keyword);=0A=
    } else {=0A=
		alert("Please input a keyword for ADS searching before clicking go");=0A=
		return false;=0A=
    }=0A=
}=0A=
/*=0A=
function processADSSearch(hiddenForm ,queryForm , str) {=0A=
    var terms =3D str;=0A=
    var formToSubmit =3D hiddenForm;=0A=
    var formToProcess =3D queryForm;=0A=
    formToSubmit.qsearch.value =3D terms;=0A=
    =
window.open("","popup","toolbar=3Dyes,location=3Dyes,status=3Dyes,menubar=
=3Dno,scrollbars=3Dyes,resizable=3Dyes,width=3D800,height=3D500");=0A=
	formToSubmit.action =3D =
"http://adsabs.harvard.edu/cgi-bin/nph-basic_connect"=0A=
	formToSubmit.target =3D "popup";=0A=
    formToSubmit.submit();=0A=
}=0A=
*/=0A=
=0A=
/*=0A=
 * Process the allContentSearch form=0A=
 * ---------------------------------=0A=
 * 1. 	Perform form validation, checking for empty fields.=0A=
 *		Give the user a relevent message if required.=0A=
 *=0A=
 * 2.	Process/Assign the appropriate form elements and submit the form.=0A=
 */=0A=
function processallContentSearch(hiddenForm, queryForm)=0A=
{=0A=
	var formToSubmit =3D hiddenForm;=0A=
	var formToProcess =3D queryForm;=0A=
	=0A=
	// Do some basic form validation=0A=
	if(formToProcess.terms.value !=3D "")=0A=
	{=0A=
		formToSubmit.WISsearch1.value =3D formToProcess.terms.value;=0A=
		formToSubmit.submit();		=0A=
	}=0A=
	else=0A=
	{=0A=
		alert("You must enter a search term in the text box before you click =
go.");=0A=
	}=0A=
}=0A=
=0A=
/*=0A=
 * Process the Title Search form=0A=
 * -----------------------------=0A=
 * 1. 	Perform form validation, checking for empty fields.=0A=
 *		Give the user a relevent message if required.=0A=
 *=0A=
 * 2.	Process/Assign the appropriate form elements and submit the form.=0A=
 */=0A=
=0A=
function processTitleSearch(hiddenForm, queryForm)=0A=
{=0A=
	var formToSubmit =3D hiddenForm;=0A=
	var formToProcess =3D queryForm;=0A=
	=0A=
	// Do some basic form validation=0A=
	if(formToProcess.terms.value !=3D "")=0A=
	{=0A=
		formToSubmit.query.value =3D formToProcess.terms.value;=0A=
		formToSubmit.submit();=0A=
	}=0A=
	else=0A=
	{=0A=
		alert("You must enter a search term in the text box before you click =
go.");=0A=
	}=0A=
}=0A=
=0A=
function doCitationSearch()=0A=
{=0A=
	var form =3D document.citsearchform;=0A=
	if(checkCitationSearch())=0A=
	{=0A=
		form.action =3D "/search/allsearch";=0A=
		form.submit();=0A=
	}=0A=
}=0A=
=0A=
// Preprocesses the arguments sent to CitationSearch and checks validity =
of user inputs.=0A=
function checkCitationSearch()=0A=
{=0A=
	var supplementFound =3D 0; //increment if found;=0A=
	var hyphenFound =3D 0; //increment if found;=0A=
	var fm =3D document.citsearchform;=0A=
	var errmsg =3D "Please use the valid digits '0' to '9'";=0A=
	// remove all spaces=0A=
	fm.volume.value =3D fm.volume.value.replace(/ /g, "");=0A=
	fm.issue.value =3D fm.issue.value.replace(/ /g, "");=0A=
	fm.pages.value =3D fm.pages.value.replace(/ /g, "");=0A=
=0A=
	if (fm.issn.value.length > 0 && fm.volume.value.length > 0 && =
fm.pages.value.length > 0)=0A=
	{=0A=
		// check page range=0A=
		var pageRange =3D fm.pages.value;=0A=
		var volume =3D fm.volume.value;=0A=
		var issue =3D fm.issue.value;=0A=
		var continueFlag =3D 0;=0A=
		//check the volume=0A=
//		if (isNaN(volume))=0A=
//		{=0A=
//			// volume failed checking=0A=
//			alert("Volume is invalid. "+errmsg+".");=0A=
//			return false;=0A=
//		}=0A=
		//check the issue=0A=
		if ((issue.length !=3D 0) && isNaN(issue))=0A=
		{=0A=
			// issue failed checking=0A=
			alert("Issue is invalid. "+errmsg+" or leave blank.");=0A=
			return false;=0A=
		}=0A=
		//check the page range=0A=
		if (isNaN(pageRange))=0A=
		{=0A=
			var isValid =3D false;=0A=
			// check for hyphen=0A=
			var hyphenIndex =3D pageRange.indexOf("-");=0A=
			if ((hyphenIndex !=3D -1) && (hyphenIndex =3D=3D =
pageRange.lastIndexOf("-")))=0A=
			{=0A=
				// we have only one hyphen, so extract each number=0A=
				var firstNum  =3D pageRange.substring(0, hyphenIndex);=0A=
				var secondNum =3D pageRange.substring(hyphenIndex + 1);=0A=
				// check for S=0A=
				var firstS  =3D firstNum.indexOf("S");=0A=
				var secondS =3D secondNum.indexOf("S");=0A=
				if (firstS =3D=3D secondS)=0A=
				{=0A=
					if ((firstS =3D=3D -1) && isNum(firstNum) && isNum(secondNum) && =
(firstNum.length > 0) && (secondNum.length > 0))=0A=
						isValid =3D true;  // both inputs are digits=0A=
					else if (firstS =3D=3D 0)=0A=
					{=0A=
						var firstNumNoS  =3D firstNum.substring(1);=0A=
						var secondNumNoS =3D secondNum.substring(1);=0A=
						if (isNum(firstNumNoS) && isNum(secondNumNoS) && =
(firstNumNoS.length > 0) && (secondNumNoS.length > 0))=0A=
							isValid =3D true; // both inputs are Sdigits=0A=
					}=0A=
=0A=
				}=0A=
			}=0A=
			if (isValid)=0A=
			{=0A=
				return true;=0A=
			}=0A=
			else=0A=
			{=0A=
				alert("The page range is invalid. "+errmsg+" and a hyphen '-' if =
required.");=0A=
				return false;=0A=
			}=0A=
		}=0A=
		return true;=0A=
	}=0A=
	else=0A=
	{=0A=
		alert("Volume and pages are required fields. Please make sure data has =
been entered for both fields.");=0A=
		return false;=0A=
	}=0A=
}=0A=
=0A=
// returns true if a is an Integer.=0A=
function isNum(a)=0A=
{ return !(isNaN(a)); }=0A=
=0A=
function fixInlineLists() {=0A=
	if (!document.getElementById) return;=0A=
	var allLIs =3D document.getElementsByTagName('li');=0A=
	var bulletNode =3D document.createElement('img');=0A=
	bulletNode.src =3D"/images/icon.bullet.gif";=0A=
=0A=
	for (var i=3D0; i<allLIs.length; i++) {=0A=
		if(allLIs[i].parentNode.className =3D=3D "inline") {=0A=
			=
allLIs[i].insertBefore(bulletNode.cloneNode(true),allLIs[i].firstChild);=0A=
		}=0A=
	}=0A=
};=0A=
=0A=
=0A=
function highlightMenuItem(oid)=0A=
{=0A=
	var loc =3D new String(document.location);=0A=
	var startIndex =3D loc.lastIndexOf("/") + 1;=0A=
	var endIndex =3D loc.length;=0A=
	var page =3D loc.substring(startIndex, endIndex);=0A=
=0A=
	var links;=0A=
	if(page =3D=3D "REFERENCES")=0A=
	{=0A=
		links =3D '<a href=3D"/cgi-bin/abstract/' + oid + =
'/ABSTRACT">Abstract</a> &nbsp;|&nbsp; <strong>References</strong>';=0A=
	}=0A=
	else if (page =3D=3D "ABSTRACT" || (page.indexOf("allsearch") !=3D -1))=0A=
	{=0A=
		links =3D '<strong>Abstract</strong> &nbsp;|&nbsp; <a =
href=3D"/cgi-bin/abstract/' + oid + '/REFERENCES">References</a>';=0A=
	}=0A=
	//alert("URL: " + loc + "\nStart Index: " + startIndex + "\nEnd Index: =
" + endIndex + "\nPage: " + page + "\nLink Text: " + links);=0A=
	return links;=0A=
}=0A=
=0A=
=0A=
function selectAll(fm)=0A=
{=0A=
	//var fm =3D document.viewSelectedForm;=0A=
=0A=
	for (var i =3D 0; i < fm.elements.length; i++)=0A=
	{=0A=
		if (fm.elements[i].type =3D=3D 'checkbox' /*&& fm.elements[i].name =
=3D=3D 'ID'*/)=0A=
		{=0A=
			fm.elements[i].checked =3D true;=0A=
		}=0A=
	}=0A=
}=0A=
=0A=
=0A=
function do_popup(file)=0A=
{=0A=
		var w =3D window.open(file, 'popWin', =
'width=3D350,height=3D500,scrollbars=3Dyes');=0A=
=0A=
}=0A=
=0A=
=0A=
function mailTool(itemID, useurl, skin)=0A=
{=0A=
	//var url =3D "/cgi-bin/mailform?item=3D"+itemID;=0A=
	var url =3D "/cgi-bin/mailform?item=3D" + itemID + "&skin=3D" + skin;=0A=
	var parameters =3D =
'status=3Dno,scrollbars=3Dno,resizable=3Dyes,width=3D400,height=3D475';=0A=
=0A=
	if (useurl !=3D "")=0A=
	{=0A=
		url +=3D "&useurl=3D"+useurl;=0A=
	}=0A=
=0A=
	if ((isNaN(parseInt(itemID))=3D=3Dtrue)||(arguments.length >=3D 4 && =
arguments[3] =3D=3D 'cluster'))=0A=
	{=0A=
		url +=3D '&type=3Dcluster';=0A=
	}=0A=
=0A=
    //check whether the last 2 arguments is used for urllink.=0A=
    // urllink is needed to open up mail form for mrw pages on tomcat =
port=0A=
    if (arguments.length>=3D5 && =
arguments[arguments.length-2]=3D=3D'urllink' && =
arguments[arguments.length-1]!=3D'')=0A=
    {=0A=
        url =3D arguments[arguments.length-1] + url;=0A=
    }=0A=
=0A=
    if (arguments.length>=3D5 && arguments[3]=3D=3D'currentlink' && =
arguments[4]!=3D'')=0A=
    {=0A=
        url =3D url + '&currentLink=3D' + arguments[4];=0A=
=0A=
        //processing specific cases=0A=
        if (arguments[4]=3D=3D'jcatalogjsphdr')=0A=
        {=0A=
            //get the issn and pass it the next stage for processing=0A=
            var currloc =3D location.href;=0A=
            var issnloc =3D currloc.indexOf("&issn=3D");=0A=
            if (issnloc !=3D -1)=0A=
            {=0A=
                url =3D url + '&currentISSN=3D' + =
currloc.substring(issnloc+6);=0A=
            }=0A=
        }=0A=
        // issn is needed for cocoon rendered mrw to form the final url.=0A=
        else if ( (arguments[4]=3D=3D'cocoonmrwhome') || =
(arguments[4]=3D=3D'cocoonmrwarticle') )=0A=
        {=0A=
            if (arguments.length>=3D7 && arguments[5]=3D=3D'issn' && =
arguments[6]!=3D'')=0A=
            {=0A=
                url =3D url + '&currentISSN=3D' + arguments[6];=0A=
            }=0A=
            //getting the parameters of the currently shown mrw from the =
url=0A=
            //the method used here is a little crude, it extracts the =
values from the current url of the page the user is in =0A=
            //to pass it to the background program to reconstruct the =
requred url=0A=
            if (arguments[4]=3D=3D'cocoonmrwhome')=0A=
            {=0A=
                var currloc =3D location.href;=0A=
                var mrwParamArr =3D currloc.split("/");=0A=
                if (mrwParamArr.length >=3D 7)=0A=
                {=0A=
                    url =3D url + '&mrwPath=3D' + mrwParamArr[5];=0A=
                    var mrwFileParam =3D mrwParamArr[6];=0A=
                    var seploc =3D mrwFileParam.indexOf("#");=0A=
                    if (seploc !=3D -1)=0A=
                    {=0A=
                        url =3D url + '&mrwSection=3D' + =
mrwFileParam.substring(seploc+1);=0A=
                        mrwFileParam =3D =
mrwFileParam.substring(0,seploc);=0A=
                    }=0A=
                    seploc =3D mrwFileParam.indexOf("?");=0A=
                    if (seploc !=3D -1)=0A=
                    {=0A=
                        url =3D url + '&mrwUrlParam=3D' + =
mrwFileParam.substring(seploc+1);=0A=
                        url =3D url + '&mrwDocType=3D' + =
mrwFileParam.substring(0,seploc);=0A=
                    }=0A=
                    else=0A=
                    {=0A=
                        url =3D url + '&mrwDocType=3D' + mrwFileParam;=0A=
                    }=0A=
                }=0A=
                =0A=
            }=0A=
            else if (arguments[4]=3D=3D'cocoonmrwarticle')=0A=
            {=0A=
                var currloc =3D location.href;=0A=
                var mrwParamArr =3D currloc.split("/");=0A=
                if (mrwParamArr.length >=3D 7)=0A=
                {=0A=
                    url =3D url + '&mrwPath=3D' + mrwParamArr[5];=0A=
                    url =3D url + '&mrwAid=3D' + mrwParamArr[7];=0A=
                    url =3D url + '&mrwVersion=3D' + mrwParamArr[8];=0A=
                    var mrwFileParam =3D mrwParamArr[9];=0A=
                    var seploc =3D mrwFileParam.indexOf("#");=0A=
                    if (seploc !=3D -1)=0A=
                    {=0A=
                        url =3D url + '&mrwSection=3D' + =
mrwFileParam.substring(seploc+1);=0A=
                        url =3D url + '&mrwDocType=3D' + =
mrwFileParam.substring(0,seploc);=0A=
                    }=0A=
                    else=0A=
                    {=0A=
                        url =3D url + '&mrwDocType=3D' + mrwFileParam;=0A=
                    }=0A=
                }=0A=
            }=0A=
        }=0A=
    }=0A=
    =0A=
    //checking if a html file exists in the url=0A=
    var tmpfname =3D location.href;=0A=
    var loc =3D tmpfname.indexOf("?");=0A=
    if (loc !=3D -1)=0A=
    {=0A=
        tmpfname =3D tmpfname.substring(0,loc);=0A=
    }=0A=
    loc =3D tmpfname.indexOf(".html");=0A=
    if (loc !=3D -1)=0A=
    {=0A=
        loc =3D tmpfname.lastIndexOf("/");=0A=
        if (loc !=3D -1)=0A=
        {=0A=
            tmpfname =3D tmpfname.substring(loc+1);=0A=
            url =3D url + '&currentFile=3D' + tmpfname;=0A=
        }=0A=
    }=0A=
=0A=
    if (arguments.length>=3D7 && arguments[5]=3D=3D'currentyear' && =
arguments[6]!=3D'')=0A=
    {=0A=
        url =3D url + '&currentYear=3D' + arguments[6];=0A=
    }=0A=
=0A=
	eval("wileyMailTool =3D =
window.open('"+url+"','MailTool','"+parameters+"')");=0A=
=0A=
	// If object exists ensure has focus=0A=
	if (eval("wileyMailTool") && window.focus)=0A=
		eval("wileyMailTool").focus();=0A=
}=0A=
=0A=
=0A=
/*=0A=
 *	1.	Find the links we need to alter.=0A=
 * 	2.	For each link apply the new styles we need.=0A=
 */=0A=
function fixAbsRefRendering()=0A=
{	=0A=
	var table =3D document.getElementById('prerendered');=0A=
	var anchors =3D table.getElementsByTagName('a');=0A=
	for (var i =3D 1; i < anchors.length; i++)=0A=
	{=0A=
		if(anchors[i].href !=3D"") =0A=
		{		=0A=
			applyAbsRefStyleCorrection(anchors[i]);=0A=
			=0A=
		}=0A=
	}=0A=
}=0A=
=0A=
function applyAbsRefStyleCorrection(currentLink)=0A=
{=0A=
	currentLink.style["color"] =3D "#369";=0A=
	currentLink.style["fontWeight"] =3D "normal";=0A=
	currentLink.style["marginLeft"] =3D "3px";=0A=
	currentLink.style["marginRight"] =3D "5px";=0A=
}=0A=
=0A=
function populatePage(formName)=0A=
{=0A=
	if (formName.Page.value =3D=3D '')=0A=
	{=0A=
		formName.Page.value =3D document.location;=0A=
	}=0A=
}=0A=
=0A=
function equalizeColumnWidth(numFeatures)=0A=
{=0A=
	var numberOfFeatures =3D numFeatures;		=0A=
	// Percentage width of table to use for each column.=0A=
	var percentPerFeature =3D Math.floor(100 / numberOfFeatures);	=0A=
	// Number of columns in total. Content cols + seperators.	=0A=
	=0A=
	document.writeln("<tr>");=0A=
	for(var i =3D 0; i < numberOfFeatures; i++)=0A=
	{		=0A=
		document.writeln('<td width=3D"' + percentPerFeature + '%"><img =
src=3D"/images/dot.clear.gif" width=3D"100%" height=3D"1" border=3D"0" =
alt=3D"blank" /></td>');=0A=
	}=0A=
	document.writeln("</tr>");=0A=
}=0A=
=0A=
function old_equalizeColumnWidth(numFeatures)=0A=
{=0A=
	var numberOfFeatures =3D numFeatures;		=0A=
	// Percentage width of table to use for each column.=0A=
	var percentPerFeature =3D Math.floor(100 / numberOfFeatures);	=0A=
	// Number of columns in total. Content cols + seperators.	=0A=
	=0A=
	if(debug)=0A=
	{=0A=
		alert("Number of feature boxes: " + numberOfFeatures + "\nPercent per =
feature: " + percentPerFeature);=0A=
	}=0A=
	=0A=
	document.writeln("<tr>");=0A=
	for(var i =3D 0; i < numberOfFeatures; i++)=0A=
	{		=0A=
		if (netscape4 =3D=3D false)=0A=
		{=0A=
			document.writeln('<td style=3D"width: ' + percentPerFeature + =
'%"><img src=3D"/images/dot.clear.gif" width=3D"1" height=3D"1" =
border=3D"0" alt=3D"blank" /></td>');=0A=
		}=0A=
		else // protect against inline style application hanging NN4.=0A=
		{=0A=
			document.writeln('<td><img src=3D"/images/dot.clear.gif" width=3D"1" =
height=3D"1" border=3D"0" alt=3D"blank" /></td>');		=0A=
		}=0A=
	}=0A=
	document.writeln("<tr>");=0A=
}=0A=
=0A=
function writeFBoxCell()=0A=
{=0A=
	if(first)=0A=
	{=0A=
		document.write('<td valign=3D"top" id=3D"firstWideFBoxCell">');=0A=
	}else=0A=
	{=0A=
		document.write('<td valign=3D"top" id=3D"wideFBoxCell">');=0A=
	}=0A=
	numFeatures ++;=0A=
}=0A=
=0A=
=0A=
//=0A=
// escape apostrophe characters within a string=0A=
//=0A=
function encode(str)=0A=
{=0A=
        out =3D '';=0A=
=0A=
        for (a =3D 0; a < str.length; a++)=0A=
        {=0A=
                c =3D str.charAt(a);=0A=
=0A=
                if (c =3D=3D "'")=0A=
                {=0A=
                        out +=3D '\\';=0A=
                }=0A=
=0A=
                out +=3D c;=0A=
        }=0A=
=0A=
        return out;=0A=
}=0A=
=0A=
=0A=
//=0A=
//=0A=
//=0A=
function escapeEntities(str)=0A=
{=0A=
	while (str.indexOf("&apos;") > -1)=0A=
	{=0A=
		pos =3D str.indexOf("&apos;");=0A=
		str =3D str.substr(0, pos) + '\\\'' + str.substr(pos + 6);=0A=
	}=0A=
=0A=
	return str;=0A=
}=0A=
=0A=
=0A=
// new popup function less intrusive just uses the DOM to pickup links =
with an exwin class.=0A=
// currently implemented for Figure popups on D&T homepage.=0A=
=0A=
function exLinks()=0A=
{=0A=
	if (document.getElementById)=0A=
	{=0A=
    	var aObject =3D document.getElementsByTagName("a");=0A=
    	for (var x=3D0; x<aObject.length; x++)=0A=
    	{				=0A=
    		if (aObject[x].className =3D=3D "exwin")=0A=
    		{=0A=
    			aObject[x].onclick =3D function()=0A=
    			{=0A=
    				=0A=
    				//if we direct through "jabout" the page will get templated, we =
dont want that so we're gonna use "hompages"=0A=
    				this.href =3D =
String(this.href).toLowerCase().replace("cgi-bin/jabout", "homepages");=0A=
    				window.open(this.href, "Figures", =
"dependent=3D0,toolbar=3D1,location=3D0,status=3D0,menubar=3D0,scrollbars=
=3D1,resizable=3D1,width=3D600,height=3D400");=0A=
    				return false;=0A=
    			}=0A=
    		}=0A=
    	}=0A=
	}=0A=
}=0A=
=0A=
=0A=
/*=0A=
* Andy Perry=0A=
* JavaScript enhancements=0A=
*/=0A=
=0A=
// One variable to act as global container for all js activity=0A=
if(typeof WIS=3D=3D"undefined") {=0A=
	var WIS=3D{=0A=
		author:"Andy Perry",=0A=
		creationDate:"31/08/2007"=0A=
	};=0A=
};=0A=
=0A=
WIS.FunctionStack =3D function() {=0A=
	this.stackArray=3D[];	=0A=
};=0A=
=0A=
WIS.FunctionStack.prototype =3D {=0A=
	addCall:function(func) {=0A=
		this.stackArray[this.stackArray.length]=3Dfunc;=0A=
	},=0A=
	executeStackCalls:function() {=0A=
		for(var i=3D0; i<this.stackArray.length; i++) {=0A=
			this.stackArray[i]();=0A=
			this.stackArray.splice(i--,1);=0A=
		}=0A=
	}=0A=
};=0A=
=0A=
if(typeof WIS.debug=3D=3D"undefined") {=0A=
	WIS.debug=3D{};=0A=
}; =0A=
=0A=
WIS.debug.XslDebug=3Dfunction() {=0A=
	var allElementsColl=3D[];=0A=
=0A=
	var allColl=3Ddocument.getElementsByTagName("*");=0A=
	for(var i=3D0;i<allColl.length;i++) {=0A=
		if(allColl[i].getAttribute("xsldebug")!=3Dnull) {=0A=
			allColl[i].style.border=3D"3px solid red";=0A=
			allColl[i].style.position=3D"relative";=0A=
			var labelDiv=3Ddocument.createElement("DIV");=0A=
			allColl[i].appendChild(labelDiv);=0A=
			var labelDivStyles=3D{backgroundColor:"#999999",border:"2px solid =
yellow",position:"absolute",top:"0",left:"0",color:"#000000"};=0A=
			for(a in labelDivStyles) {=0A=
				labelDiv.style[a]=3DlabelDivStyles[a];=0A=
			};=0A=
			labelDiv.innerHTML=3D"something: =
"+allColl[i].getAttribute("xsldebug").substr(0,allColl[i].getAttribute("x=
sldebug").indexOf("::"));=0A=
		}=0A=
	}=0A=
}=0A=
=0A=
=0A=
function addTitle(oid,salesModelId) {=0A=
	http.open("POST", =
"/cart/managesubscription?oid=3D"+oid+"&action=3Dadd&displayCart=3Dfalse&=
salesModelId=3D"+salesModelId, true); =0A=
	=0A=
	http.onreadystatechange=3Dfunction() {=0A=
		if(http.readyState =3D=3D 4) {=0A=
			document.getElementById('cart').innerHTML=3D'Title added to My Cart'; =0A=
	    	toggleShCrtLink();=0A=
	    }=0A=
	}=0A=
	http.send(null);=0A=
	=0A=
};=0A=
=0A=

------=_NextPart_000_0000_01C99F0E.BB8D06B0
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www3.interscience.wiley.com/freeflow/js/menu.js

/*=0A=
* Project				:	Interscience Accessibility=0A=
* File					:	menu.js=0A=
* Creation Date				:	26th May 2005=0A=
* Author				:	Ben Marvell=0A=
*=0A=
* Various functions to deliver personalisation functionality.=0A=
*/=0A=
=0A=
// route browser to required menu function=0A=
var runonce =3D false;=0A=
var dom =3D (document.getElementById) ? true : false;=0A=
=0A=
var rollimg =3D new Image();=0A=
rollimg.src =3D =
"http://download.interscience.wiley.com/images/menu.arrow.1.gif";=0A=
=0A=
function wisMenu(){=0A=
	if (!runonce){=0A=
		runonce =3D true;=0A=
		if ((navigator.userAgent.indexOf("Mac") !=3D -1) && =
(navigator.userAgent.indexOf("MSIE") !=3D -1) || =
(navigator.userAgent.indexOf("Netscape6") !=3D -1)){=0A=
			return;=0A=
		} else if (dom){=0A=
			var classname =3D "menuDOM";=0A=
			menuHoverDOM(classname);=0A=
			fixInlineLists(); // fix the inline lists=0A=
		}=0A=
	}else{=0A=
		if ((navigator.userAgent.indexOf("Mac") !=3D -1) && =
(navigator.userAgent.indexOf("MSIE") !=3D -1) || =
(navigator.userAgent.indexOf("Netscape6") !=3D -1)){=0A=
		    var classname =3D "menuIEMAC";=0A=
		    menuHoverDOM(classname);=0A=
		} else if (dom){=0A=
			fixInlineLists(); // fix the inline lists=0A=
			return;=0A=
		} else if(document.all){=0A=
			fixMenuIE4();=0A=
		}=0A=
	}=0A=
}=0A=
=0A=
// dom supported browsers=0A=
//@@@DM It was anoying me this error=0A=
var menuHoverDOM =3D function(classname){=0A=
	=0A=
	var subnodeslength =3D (typeof(submenulength) !=3D "undefined") ? =
(submenulength + 1) : 8;=0A=
	if (document.getElementById("menu")){=0A=
		var menu =3D =
document.getElementById("menu").getElementsByTagName("UL")[0];=0A=
		=0A=
		//	AP	06/12/2007	BUG3622	- Prevent error on production interface if =
personalisation bar doesn't exist.=0A=
		if(typeof menu =3D=3D "undefined") return;=0A=
		=0A=
		// apply menuDOM class and user class=0A=
		menu.className =3D classname;=0A=
		if (document.getElementById("user")){=0A=
			document.getElementById("user").className =3D classname;=0A=
		}=0A=
		//if no userdata loaded then padd the lists to the right=0A=
		if (menu.getElementsByTagName("LI").length =3D=3D 2){=0A=
			menu.style.paddingLeft =3D "5em";=0A=
		}=0A=
		=0A=
		//loop through li elements=0A=
		for (var x=3D0; x<menu.childNodes.length; x++){=0A=
		    if (menu.childNodes[x].nodeName =3D=3D "LI"){	      =0A=
		        for (var y=3D0; y<menu.childNodes[x].childNodes.length; y++){  =
      =0A=
		            //if element has sub menu apply appropirate image class =
(ie all code in this statment is for drop menus)=0A=
		            if (menu.childNodes[x].childNodes[y].nodeName =3D=3D "UL"){=0A=
		            	// setup some common variables to make the code shorter		 =
       =0A=
		            	var lilength =3D =
parseInt(menu.childNodes[x].childNodes[y].getElementsByTagName("LI").leng=
th);		            =0A=
		            	var liels =3D =
menu.childNodes[x].childNodes[y].getElementsByTagName("LI");=0A=
		            	// set parent li to have the sub menu arrow=0A=
		                menu.childNodes[x].className =3D "domArrowOut";=0A=
		               		               		                =0A=
		                // set the amount of saved titles for the parent li's =
ie (5 saved searches)=0A=
		                var menuamount =3D (lilength-3);=0A=
		                if (environment =3D=3D 'PRODUCTION') {=0A=
		                	menuamount =3D (lilength-2);=0A=
		                }=0A=
		                if (menuamount < 0) menuamount =3D 0;=0A=
		                =
menu.childNodes[x].getElementsByTagName("A")[0].innerHTML =3D menuamount =
+ " " + menu.childNodes[x].getElementsByTagName("A")[0].innerHTML;=0A=
		                =0A=
		                // checks the submenu and shows no data message if =
user doesnt have any data saved		                		              =0A=
	                    var tmpinnerhtml =3D "";=0A=
	                    var minLength =3D 3;=0A=
	                    if (environment =3D=3D 'PRODUCTION') {=0A=
	                    	minLength =3D 2;=0A=
	                    }=0A=
	                    if (lilength <=3D minLength){	                 =0A=
		                    for (var j=3D0; j<lilength; j++){                  =
        		                       =0A=
	                            if (j=3D=3D0)=0A=
	                                tmpinnerhtml +=3D "<li>" + =
liels[j].innerHTML + "</li><li class=3D'nosaved'><em>You do not have =
anything saved in this menu.</em></li>";=0A=
	                            else=0A=
	                                tmpinnerhtml +=3D "<li class=3D'hide'>" =
+ liels[j].innerHTML + "</li>";	                            =0A=
	                    	}	                    =0A=
	                    } else {=0A=
	                    	//setup var to see if more is required=0A=
	                    	var more =3D (subnodeslength < lilength) ? true : =
false;	                    	=0A=
	                    	var flag =3D true;=0A=
	                    	for (var j=3D0; j<lilength; j++){	                 =
   	=0A=
	                    		// if we only have 8 menu elements then just =
display them else delete the rest and add a more button=0A=
	                    		if (j < subnodeslength){=0A=
	                    			tmpinnerhtml +=3D "<li>" + liels[j].innerHTML + =
"</li>";=0A=
	                    		} else if (j > lilength-3){=0A=
	                    			if (j =3D=3D lilength-2){=0A=
	                    				var url =3D liels[j].innerHTML.split('"')[1];=0A=
	                    				tmpinnerhtml +=3D "<li><a href=3D'"+url+"' =
target=3D'_top'>more...</a></li><li>" + liels[j].innerHTML + "</li>";=0A=
	                    			} else {=0A=
	                    				tmpinnerhtml +=3D "<li>" + liels[j].innerHTML + =
"</li>"; =0A=
	                    			}                 			=0A=
	                    		}	=0A=
	                    	}=0A=
	                    	// set last two menu items to have no rollover and =
a border at the top                   	=0A=
	                    }=0A=
	                    menu.childNodes[x].childNodes[y].innerHTML =3D =
tmpinnerhtml;=0A=
	                    // set default menu elements with diff style=0A=
	                    if (flag){=0A=
		                    if (more){=0A=
				                liels[liels.length-3].style.borderTop =3D "1px solid =
#DDDDDD";=0A=
				                liels[liels.length-3].className =3D "noroll";=0A=
				                liels[liels.length-2].style.borderTop =3D "1px solid =
#DDDDDD";=0A=
				                liels[liels.length-2].className =3D "noroll";=0A=
				                liels[liels.length-1].className =3D "noroll";=0A=
				                more =3D false;=0A=
					        } else {=0A=
				            	liels[liels.length-2].style.borderTop =3D "1px solid =
#DDDDDD";=0A=
				                liels[liels.length-2].className =3D "noroll";=0A=
				                liels[liels.length-1].className =3D "noroll";=0A=
					        }					        =0A=
							flag =3D false;					     =0A=
					    } =0A=
	                    	                    =0A=
		                // set sub menu title for the msnu=0A=
		                liels[0].className =3D "hoverTitle";=0A=
		                // apply rollover functions to hide/show sub menus=0A=
		                if (classname =3D=3D "menuDOM"){=0A=
	    	                menu.childNodes[x].onmouseover =3D function(){=0A=
	        	                // safari doesnt handle auto in absoulte =
positioning correctly so this code makes the menus display correctly.=0A=
								if (navigator.userAgent.indexOf("Safari") !=3D -1){=0A=
								   	if(!window.devicePixelRatio) {=0A=
									 this.getElementsByTagName("UL")[0].style.marginTop =3D "20px";=0A=
									 this.getElementsByTagName("UL")[0].style.marginLeft =3D "0";=0A=
								  	}=0A=
								}=0A=
	        	                this.className =3D"menuhover domArrowOver";=0A=
								if(document.all) =
this.getElementsByTagName("iframe")[0].style.height=3Dthis.getElementsByT=
agName("ul")[0].offsetHeight;=0A=
	        	            }=0A=
	        	            menu.childNodes[x].onmouseout=3Dfunction() {=0A=
	            			    this.className =3D "domArrowOut";=0A=
	        		        }=0A=
	    		        }=0A=
=0A=
						if(document.all) {=0A=
							var newIframe =3D document.createElement("iframe");=0A=
							newIframe.style.filter =3D =
"progid:DXImageTransform.Microsoft.Alpha(style=3D0,opacity=3D0)";=0A=
							menu.childNodes[x].appendChild(newIframe);=0A=
						}=0A=
=0A=
		            }            =0A=
		        }=0A=
		        // deletes the sub menus on the IEMAC as they have a =
background bug, will still remain on NS6=0A=
		    	if ((navigator.userAgent.indexOf("Mac") !=3D -1) && =
(navigator.userAgent.indexOf("MSIE") !=3D -1)){=0A=
	            	menu.childNodes[x].innerHTML =3D =
menu.childNodes[x].innerHTML.split("<UL")[0];=0A=
	            }                    	=0A=
		    }=0A=
		}=0A=
	}=0A=
};=0A=
//@@@DM It was anoying me this error		=0A=
var fixMenuIE4 =3D function(){=0A=
	if (document.all["menu"]){=0A=
		var tmptable =3D "<table class=3D'menuIEFOUR' align=3D'right' =
border=3D'0' cellpadding=3D'0' cellspacing=3D'0' height=3D'25'><tr>"=0A=
		var menu =3D document.all["menu"].children[0];=0A=
		menu.style.marginBottom =3D "-3px";=0A=
		for (var x=3D1; x<menu.children.length; x++){=0A=
	        if (userName !=3D ""){=0A=
		        var tmp =3D menu.children[x].innerHTML.split("<UL")[0];=0A=
				if (x=3D=3D1){=0A=
				    tmptable +=3D "<td width=3D'15'></td><td width=3D'180' =
class=3D'menuIEFOURUser'><span>" + tmp + "</span></td>";=0A=
				} else if (x=3D=3D5 || x=3D=3D6){ =0A=
				    tmptable +=3D "<td width=3D'15' =
class=3D'menuIEFOURDOWN'></td><td width=3D'70'><span>" + tmp + =
"</span></td>";=0A=
				} else {=0A=
				    tmptable +=3D "<td width=3D'15' =
class=3D'menuIEFOURSIDE'></td><td width=3D'70'><span>" + tmp + =
"</span></td>";=0A=
				}=0A=
				if (x =3D=3D (menu.children.length-1)){=0A=
					tmptable +=3D "</tr></table>";	=0A=
				}=0A=
			} else {=0A=
				var tmp =3D menu.children[x].innerHTML.split("<UL")[0];=0A=
				if (x=3D=3D1){ 				 =0A=
					tmptable +=3D "<td width=3D'40'></td><td width=3D'15' =
class=3D'menuIEFOURDOWN'></td><td width=3D'70'><span>" + tmp + =
"</span></td>";=0A=
				} else if (x=3D=3D2) {=0A=
				    tmptable +=3D "<td width=3D'15' =
class=3D'menuIEFOURDOWN'></td><td width=3D'50'><span>" + tmp + =
"</span></td></tr></table>";=0A=
				}=0A=
			}=0A=
		}=0A=
		menu.innerHTML =3D tmptable;=0A=
	}=0A=
};=0A=
=0A=
var menuloadedflag =3D true;=0A=
=0A=
=0A=
var time =3D new Date();=0A=
//@@@DM It was anoying me this error		=0A=
var ordval=3D (time.getTime());=0A=

------=_NextPart_000_0000_01C99F0E.BB8D06B0
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www3.interscience.wiley.com/freeflow/js/journalhome.js

//=0A=
// Homepage links format:=0A=
// homepagelinks[i][0] =3D location=0A=
// homepagelinks[i][1] =3D currenthref=0A=
// homepagelinks[i][2] =3D type: "e" =3D end of line, "s" =3D separator, =
"" =3D normal=0A=
//=0A=
var writeLinksTraceOn =3D false;=0A=
function writeJournalLinks(filename, oid)=0A=
{=0A=
	var url =3D new String(document.location);=0A=
	var urlpathinfo  =3D "";=0A=
	var pathinfomatch =3D false;=0A=
=0A=
	// Find the pathinfo in the URL - need it later (probably)=0A=
	//=0A=
	// Both url and currenthref are expected to be of the form:=0A=
	// ...../<cgi-name>/<oid>[/<pathinfo>]=0A=
	//=0A=
	=0A=
	urlindex =3D url.indexOf("jabout");=0A=
	if (urlindex =3D=3D -1)=0A=
	{=0A=
		urlindex =3D url.indexOf("mrwhome");=0A=
	}=0A=
	if (urlindex =3D=3D -1)=0A=
	{=0A=
		urlindex =3D url.indexOf("collectionhome");=0A=
	}=0A=
	if (urlindex =3D=3D -1)=0A=
	{=0A=
		urlindex =3D url.indexOf("bookhome");=0A=
	}=0A=
=0A=
	if (urlindex !=3D -1)	// If found a known CGI name....=0A=
	{=0A=
		// We needs a pathinfo match.....=0A=
		pathinfomatch =3D true;=0A=
=0A=
		// Find the end of the cgi-name=0A=
		urlindex =3D url.indexOf("/", urlindex + 1);=0A=
=0A=
		// Look for the start of pathinfo=0A=
		urlindex =3D url.indexOf("/", urlindex + 1);=0A=
=0A=
		if (urlindex !=3D -1)=0A=
		{=0A=
			urlindex1 =3D url.indexOf("?", urlindex + 1);=0A=
=0A=
			if(urlindex1 !=3D -1){=0A=
				urlpathinfo =3D url.substring(urlindex + 1, urlindex1);=0A=
				writeLinksTrace("urlpathinfo: " + urlpathinfo);						=0A=
			}else{			=0A=
				urlpathinfo =3D url.substring(urlindex + 1, url.length);=0A=
				writeLinksTrace("urlpathinfo: " + urlpathinfo);=0A=
			}=0A=
		}=0A=
	}=0A=
=0A=
	var colnum =3D 0;=0A=
=0A=
	for (var i =3D 0; i < homepagelinks.length; i++)=0A=
	{=0A=
		var location    =3D homepagelinks[i][0];=0A=
		var currenthref =3D homepagelinks[i][1];=0A=
		var type        =3D homepagelinks[i][2];=0A=
=0A=
		writeLinksTrace(location + " : " + currenthref + " : " + type);=0A=
=0A=
		if(type =3D=3D "x") // End of the list=0A=
		{=0A=
			break;=0A=
		}=0A=
		if(type =3D=3D "s")=0A=
		{=0A=
			document.writeln('</span><br />');=0A=
			colnum =3D 0;=0A=
			continue;=0A=
		}=0A=
		if (colnum > 0)=0A=
		{=0A=
			document.writeln('&nbsp;|&nbsp;');=0A=
		}=0A=
=0A=
		var hrefindex;=0A=
		var urlindex;=0A=
		var greydisplay =3D false;=0A=
=0A=
		// See what type of link we have=0A=
=0A=
		// Cases here are for URLs with no /relevant/ pathinfo=0A=
		if (pathinfomatch !=3D true)=0A=
		{=0A=
			hrefindex =3D currenthref.indexOf("jhome");=0A=
			if (hrefindex !=3D -1)=0A=
			{=0A=
				urlindex =3D url.indexOf("jhome");=0A=
				if (urlindex !=3D -1)=0A=
				{=0A=
					greydisplay =3D true;=0A=
				}=0A=
			}=0A=
			if (hrefindex =3D=3D -1)	// Didn't find jhome, try jtoc=0A=
			{=0A=
				hrefindex =3D currenthref.indexOf("jtoc");=0A=
				if (hrefindex !=3D -1)=0A=
				{=0A=
					urlindex =3D url.indexOf("jtoc");=0A=
					if (urlindex !=3D -1)=0A=
					{=0A=
						greydisplay =3D true;=0A=
					}=0A=
				}=0A=
			}=0A=
			// Add more non-pathinfo cases here...=0A=
		}=0A=
=0A=
		// Cases under here are for cases with relevant pathinfo=0A=
		if (pathinfomatch =3D=3D true)=0A=
		{=0A=
			var hrefmatch =3D false;=0A=
			var hrefpathinfo =3D "";=0A=
=0A=
			writeLinksTrace("pathinfo handler");=0A=
=0A=
			// Find the pathinfo in the href=0A=
=0A=
			hrefindex =3D currenthref.indexOf("jabout");=0A=
			if (hrefindex =3D=3D -1)=0A=
			{=0A=
				hrefindex =3D currenthref.indexOf("mrwhome");=0A=
			}=0A=
			if (hrefindex =3D=3D -1)=0A=
			{=0A=
				hrefindex =3D currenthref.indexOf("collectionhome");=0A=
			}=0A=
			if (hrefindex =3D=3D -1)=0A=
			{=0A=
				hrefindex =3D currenthref.indexOf("bookhome");=0A=
			}=0A=
			if (hrefindex !=3D -1)=0A=
			{=0A=
				// We needs a pathinfo match.....=0A=
				hrefmatch =3D true;=0A=
=0A=
				// Find the end of the cgi-name=0A=
				hrefindex =3D currenthref.indexOf("/", hrefindex + 1);=0A=
=0A=
				// Look for the start of pathinfo=0A=
				hrefindex =3D currenthref.indexOf("/", hrefindex + 1);=0A=
=0A=
				if (hrefindex !=3D -1)=0A=
				{=0A=
					hrefpathinfo =3D currenthref.substring(hrefindex + 1, =
currenthref.length);=0A=
					writeLinksTrace("hrefpathinfo: " + hrefpathinfo);=0A=
				}=0A=
			}=0A=
=0A=
			// If we found a CGI matches in currenthref, and the=0A=
			// pathinfos match then grey this link=0A=
			if (hrefmatch =3D=3D true && urlpathinfo =3D=3D hrefpathinfo)=0A=
			{=0A=
				greydisplay =3D true;=0A=
			}=0A=
		}=0A=
=0A=
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------=_NextPart_000_0000_01C99F0E.BB8D06B0
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------=_NextPart_000_0000_01C99F0E.BB8D06B0
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function pagetitle() {return 'Int. J. Cancer Vol.9999, 9999 Pages: NA'; =
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function dochead(hwnd, spec) {var title =3D pagetitle(); =
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hwnd.document.write('<HTML><HEAD><TITLE>'+spec+title+'</TITLE></HEAD><BOD=
Y bgcolor=3D"#ffffff">'); }   =0A=
function doctail(hwnd) {hwnd.document.write('</BODY></HTML>'); }   =0A=
function pagehead(hwnd) {hwnd.document.write('<TABLE WIDTH=3D"600" =
BORDER=3D"0" ><TR ><TD WIDTH=3D"122" HEIGHT=3D"26" VALIGN=3D"BOTTOM" =
><IMG WIDTH=3D"122" HEIGHT=3D"165" BORDER=3D"0" ALT=3D"About this =
Journal" =
SRC=3D"http://www.interscience.wiley.com/covers/0020-7136.gif"></TD ><TD =
WIDTH=3D"351" HEIGHT=3D"26" VALIGN=3D"BOTTOM" ><P ALIGN=3D"LEFT" ><FONT =
SIZE=3D"-1" >Online ISSN: 1097-0215&nbsp;&nbsp;&nbsp;&nbsp;Print ISSN: =
0020-7136</FONT ><BR><B ><FONT SIZE=3D"+1" FACE=3D"Ariel, Helvetica" =
>International Journal of Cancer</FONT ></B ><BR><FONT SIZE=3D"-1" =
FACE=3D"Ariel, Helvetica" ><B >Volume 9999, Issue 9999, 2009.</B >  =
Pages: NA</FONT ><BR><FONT ></FONT ></P ><P ALIGN=3D"LEFT" >Copyright =
&copy; 2008 Wiley-Liss, Inc.</P ></TD ><TD WIDTH=3D"82" HEIGHT=3D"26" =
VALIGN=3D"BOTTOM" ></TD ></TR ></TABLE ><HR>'); }   =0A=
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SRC=3D"'+iname+'"><BR>'); }  =0A=
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href=3Djavascript:display_'+iname); if (size =3D=3D 1) =
hwnd.document.write('(0) target=3Dmain >View Normal Image</a> '); else =
hwnd.document.write('(1) target=3Dmain >View Magnified Image</a> '); =
hwnd.document.write('<a href=3Djavascript:imagelist() target=3Dmain =
>View List of Images</a> '); hwnd.document.write('<br><br>'); }   =0A=
function caption_fig001(hwnd) { hwnd.document.write('<B >Figure 1. </B =
>Effects of NK012 and CPT-11 on U87MG/Luc tumor xenograft. (<I >a</I >) =
Tumor volume in mice treated with CPT-11 or NK012. U87MG/Luc tumor was =
subcutaneously inoculated into the flank of mice, as described in the <A =
HREF=3D"main.html,ftx_abs#SEC1-1" TARGET=3D"main" >Materials and =
methods</A > section. NaCl (0.9%) solution (<IMG =
SRC=3D"/giflibrary/12/bull.gif" BORDER=3D"0">), CPT-11 at 66.7 mg/kg =
(<IMG SRC=3D"/giflibrary/12/squf.gif" BORDER=3D"0">) and NK012 at 30 =
mg/kg (<IMG SRC=3D"/giflibrary/12/utrif.gif" BORDER=3D"0">) were =
intravenously administered on days 0, 4 and 8 (arrows). Points, mean; =
bars, SD. *<I >p</I > &lt; 0.05. (<I >b</I >) Treatment-related body =
weight loss occurred in mice treated with CPT-11 and NK012. Points, =
mean; bars, SD.<BR><FONT SIZE=3D"-2" ></FONT >'); }  =0A=
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function caption_fig002(hwnd) { hwnd.document.write('<B >Figure 2. </B =
>Distribution of NK012 or CPT-11 in U87MG/Luc glioma xenografts. Mice =
bearing U87MG/Luc tumor were injected with NK012 (30 mg/kg/day) or =
CPT-11 (66.7 mg/kg/day). Tumor tissues were excised 2, 12 and 24 hr =
after the intravenous injection of NK012 or CPT-11. Each mouse was =
administered fluorescein-labeled <I >Lycopersicon esculentum</I > lectin =
5 min before sacrifice to detect tumor blood vessels. Frozen sections =
were examined under a fluorescence microscope at an excitation =
wavelength of 377 nm and an emission wavelength of 477 nm. The same =
fluorescence conditions can be applied for visualizing NK012 and CPT-11 =
fluorescence. Free SN-38 could not be detected under these fluorescence =
conditions. The white lines indicate the border between the tumor and =
the brain tissue. T, U87MG/Luc tumor; B, normal brain tissue. (Scale =
bars: 20 <IMG SRC=3D"/giflibrary/12/mu.gif" BORDER=3D"0" =
ALIGN=3D"ABSBOTTOM">m).<BR><FONT SIZE=3D"-2" ></FONT >'); }  =0A=
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>Plasma, brain tissue and orthotopic tumor concentrations of respective =
analytes after intravenous administration of NK012 (30 mg/kg/day) and =
CPT-11 (66.7 mg/kg/day) to U87MG/Luc-bearing nude mice. (<I >a</I >) =
plasma; (<I >b</I >) tumor. <IMG SRC=3D"/giflibrary/12/bull.gif" =
BORDER=3D"0">, polymer-bound SN-38; <IMG SRC=3D"/giflibrary/12/cir.gif" =
BORDER=3D"0">, free SN-38 (polymer-unbound SN-38); <IMG =
SRC=3D"/giflibrary/12/utrif.gif" BORDER=3D"0">, CPT-11; <IMG =
SRC=3D"/giflibrary/12/Delta.gif" BORDER=3D"0">, free SN-38 converted =
from CPT-11.<BR><FONT SIZE=3D"-2" ></FONT >'); }  =0A=
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function caption_fig004(hwnd) { hwnd.document.write('<B >Figure 4. </B =
>Antitumor effect of NK012 or CPT-11 on orthotopic xenograft and =
survival. Mice receiving intracranial injections of U87MG/Luc were =
assigned into groups 20 days after tumor inoculation. Mice were =
intravenously administered with 0.9% NaCl solution (<IMG =
SRC=3D"/giflibrary/12/bull.gif" BORDER=3D"0">), NK012 (30 mg/kg/day, =
<IMG SRC=3D"/giflibrary/12/utrif.gif" BORDER=3D"0">) or CPT-11 (66.7 =
mg/kg/day, <IMG SRC=3D"/giflibrary/12/squf.gif" BORDER=3D"0">) on days 0 =
(20 days after tumor inoculation), 4 and 8 (arrows). (<I >a</I >) =
Representative luminescence intensity images obtained in individual =
control and treatment-group mice on the days indicated. (<I >b</I >) =
Antitumor effect of NK012 or CPT-11 on days 14, 21 and 28. Each group =
consisted of 4 mice. Points, mean; bars, SD. *<I >p</I > &lt; 0.05. (<I =
>c</I >) Treatment effects of NK012 on survival. Survival was assessed =
by Kaplan-Meier analysis. Each group consisted of 6 mice. Experiments =
were repeated twice with similar results.<BR><FONT SIZE=3D"-2" ></FONT =
>'); }  =0A=
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      <DIV class=3Dprerendered><!-- Rendered from source SGML using =
DSSSL --><!-- Rendering Developed By Andy Townsend. June-99 Aug-00 =
--><FONT=20
      size=3D+1>&nbsp;Fast Track</FONT><BR>
      <TABLE width=3D"100%">
        <TBODY>
        <TR>
          <TD>
            <DIV class=3DarticleTitle>Potent antitumor effect of=20
            SN-38-incorporating polymeric micelle, NK012, against =
malignant=20
            glioma</DIV>
            <DIV class=3DarticleSubTitle></DIV></TD></TR>
        <TR>
          <TD>Jun-ichiro Kuroda<SUP><FONT size=3D-1> 1 2</FONT></SUP>, =
Jun-ichi=20
            Kuratsu<SUP><FONT size=3D-1> 2</FONT></SUP>, Masahiro=20
            Yasunaga<SUP><FONT size=3D-1> 1</FONT></SUP>, Yoshikatsu=20
            Koga<SUP><FONT size=3D-1> 1</FONT></SUP>, Yohei =
Saito<SUP><FONT=20
            size=3D-1> 1</FONT></SUP>, Yasuhiro Matsumura<SUP><FONT =
size=3D-1>=20
            1</FONT><FONT size=3D-1> *</FONT></SUP><SUP><IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/10/dagger.gif"></SUP=
></TD></TR>
        <TR>
          <TD><SUP><FONT size=3D-1>1</FONT></SUP>Investigative Treatment =

            Division, Research Center for Innovative Oncology, National =
Cancer=20
            Center Hospital East, Kashiwa, Japan<BR><SUP><FONT=20
            size=3D-1>2</FONT></SUP>Department of Neurosurgery, Faculty =
of Medical=20
            and Pharmaceutical Sciences, Kumamoto University, Kumamoto,=20
          Japan<BR></TD></TR>
        <TR>
          <TD><B>email:</B> Yasuhiro Matsumura (<A=20
            =
href=3D"mailto:yhmatsum@east.ncc.go.jp">yhmatsum@east.ncc.go.jp</A>)</TD>=
</TR></TBODY></TABLE>
      <P><SUP><FONT size=3D-1>*</FONT></SUP>Correspondence to Yasuhiro =
Matsumura,=20
      Investigative Treatment Division, Research Center for Innovative =
Oncology,=20
      National Cancer Center Hospital East, 6-5-1 Kashiwanoha, Kashiwa =
277-8577,=20
      Japan</P><SUP><IMG border=3D0=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/10/dagger.gif"></SUP=
>Fax:=20
      +81-4-7134-6857<BR>
      <SCRIPT =
language=3DJavaScript>setDOI("ADOI=3D10.1002/ijc.24171")</SCRIPT>

      <P><B>Funded by:</B><BR><IMG align=3D0=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/bull.gif">&nbsp;M=
inistry=20
      of Health<BR><IMG align=3D0=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/bull.gif">&nbsp;L=
abour=20
      and Welfare of Japan (Third Term Comprehensive Contorol Research =
for=20
      Cancer)<BR><IMG align=3D0=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/bull.gif">&nbsp;C=
ore=20
      Research for Evolutional Science and Technology<BR><IMG align=3D0=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/bull.gif">&nbsp;M=
inistry=20
      of Education, Culture, Sports, Science and Technology</P>
      <P>
      <TABLE border=3D0 width=3D"100%">
        <TBODY>
        <TR>
          <TD class=3DmainSectionHeader><A=20
        name=3Dkeywords>Keywords</A></TD></TR></TBODY></TABLE>
      <TABLE>
        <TBODY>
        <TR>
          <TD>glioma =95 drug delivery system =95 blood brain barrier =
(BBB) =95=20
            SN-38 =95 micelles</TD></TR></TBODY></TABLE>
      <P>
      <TABLE border=3D0 width=3D"100%">
        <TBODY>
        <TR>
          <TD class=3DmainSectionHeader><A=20
        name=3Dabstract>Abstract</A></TD></TR></TBODY></TABLE>
      <TABLE>
        <TBODY>
        <TR>
          <TD><IMG border=3D0 alt=3DAbstract=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_here_small.gif"></TD=
>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-1"><IMG=20
            border=3D0 alt=3D"Material and methods"=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-2"><IMG=20
            border=3D0 alt=3DResults=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-3"><IMG=20
            border=3D0 alt=3DDiscussion=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#acknowledgements"><IMG=20
            border=3D0 alt=3DAcknowledgements=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#references"><IMG=20
            border=3D0 alt=3DReferences=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD></TR></TBODY></TABLE>
      <TABLE border=3D0 width=3D"100%">
        <TBODY>
        <TR>
          <TD>Recent published reports on clinical trials of CPT-11 =
indicate=20
            the effectiveness of this compound, a prodrug of SN-38, =
against=20
            malignant glioma in combination with anti-vascular =
endothelial=20
            growth factor antibody. Here, we determined if NK012, and =
SN-38=20
            incorporating micelle, can be an appropriate formulation for =

            glioblastoma treatment compared with CPT-11. <I>In vitro</I> =

            cytotoxicity was evaluated against several glioma lines with =
NK012,=20
            CPT-11, SN-38, ACNU, CDDP and etoposide. For the <I>in =
vivo</I>=20
            test, a human glioma line (U87MG) transfected with the=20
            <I>luciferase</I> gene was inoculated into nude mice brain =
for=20
            pharmacokinetic analysis by fluorescence microscopy and=20
            high-performance liquid chromatography after intravenous =
injection=20
            of NK012 and CPT-11. <I>In vivo</I> antitumor activity of =
NK012 and=20
            CPT-11 was evaluated by bioluminescence image and =
Kaplan-Meier=20
            analyses. The growth-inhibitory effects of NK012 were 34- to =

            444-fold more potent than those of CPT-11. Markedly enhanced =
and=20
            prolonged distribution of free SN-38 in the xenografts was =
observed=20
            after NK012 injection compared with CPT-11. NK012 showed=20
            significantly potent antitumor activity against an =
orthotopic=20
            glioblastoma multiforme xenograft and significantly longer =
survival=20
            rate than CPT-11 (<I>p</I> =3D 0.0014). This implies that =
NK012 can=20
            pass through the blood brain tumor barrier effectively. =
NK012, which=20
            combines enhanced distribution with prolonged sustained =
release, may=20
            be ideal for glioma treatment. Currently, a phase I study of =
NK012=20
            is almost complete in Japan and the US. The present =
translational=20
            study warrants the clinical phase II study of NK012 in =
patients with=20
            malignant glioma. =A9 2008 Wiley-Liss, =
Inc.</TD></TR></TBODY></TABLE>
      <HR align=3Dleft SIZE=3D1 width=3D"25%">
      Received: 10 September 2008; Accepted: 28 October 2008
      <P>
      <TABLE border=3D0 width=3D"100%">
        <TBODY>
        <TR>
          <TD class=3DmainSectionHeader><A name=3Ddoi>Digital Object =
Identifier=20
            =
(DOI)</A></TD></TR></TBODY></TABLE><BR>10.1002/ijc.24171&nbsp;&nbsp;<A=20
      href=3D"http://www3.interscience.wiley.com/doiinfo.html" =
target=3Dhelp>About=20
      DOI</A><BR>
      <P>
      <TABLE border=3D0 width=3D"100%">
        <TBODY>
        <TR>
          <TD class=3DmainSectionHeader><A name=3Darticletext>Article=20
        Text</A></TD></TR></TBODY></TABLE><BR>
      <P>Malignant astrocytomas, such as anaplastic astrocytoma and =
glioblastoma=20
      multiforme (GBM), are the most common and highly vascularized =
glial tumors=20
      of the brain. At least 80 percent of malignant gliomas are =
categorized as=20
      GBM.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB1">1</A>]=20
      Currently, GBM patients have a mean survival of only 50 weeks =
following=20
      the standard treatment consisting of surgical and adjuvant =
therapies.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB2">2</A>]=20
      A recent phase III randomized trial for newly diagnosed GBM =
demonstrated=20
      that radiation therapy with concurrent temozolomide treatment, =
followed by=20
      6 months of temozolomide treatment was superior to radiation =
therapy alone=20
      in terms of overall survival.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB3">3</A>]=20
      In addition, several clinical trials have demonstrated that the =
median=20
      survival times of patients with recurrence were only 3-6 =
months.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB4">4</A>]</P>
      <P>The anticancer plant alkaloid 7-ethyl-10-hydroxy-camptothecine =
(SN-38)=20
      is a broad spectrum anticancer agent targeting DNA topoisomerase I =
with a=20
      different mechanism of action compared with alkylating agents such =
as=20
      temozolomide. Although SN-38 has shown promising anticancer =
activity <I>in=20
      vitro</I> and <I>in vivo</I>, its clinical application has =
remained=20
      dormant because of its low therapeutic efficacy and severe toxic=20
      effects.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB5">5</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB6">6</A>]=20
      Irinotecan hydrochloride (CPT-11), a prodrug of SN-38, shows some=20
      antitumor activities in patient with recurrent GBM, with response =
rates of=20
      0 to 17% in several trials.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB7">7-10</A>]=20
      CPT-11 activity is thus similar to that of other agents used for =
recurrent=20
      GBM.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB9">9</A>]=20
      A recent phase II trial for recurrent GBM demonstrated that the=20
      combination of CPT-11 and bevacizumab, an antivascular endothelial =
growth=20
      factor (VEGF) monoclonal antibody, is an effective treatment =
against the=20
      neoplasia with a 6-month progression-free survival rate of 46% and =
a=20
      6-month overall survival rate of 77%.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB11">11</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB12">12</A>]=20
      However, there is an increased risk of developing venous =
thromboembolic=20
      disease and intracranial hemorrhage with this combination therapy. =

      Therefore, there is an urgent need to develop treatment modalities =
by=20
      which cytotoxic drugs can exert more potent antitumor activity to =
their=20
      full potential with modest adverse effects and thereby reasonably =
prolong=20
      the overall survival in GBM patients.</P>
      <P>The purpose of the drug delivery system (DDS) is to achieve =
selective=20
      delivery of antitumor agents to tumor tissue at an effective =
concentration=20
      for the appropriate duration of time in order to reduce the =
adverse=20
      effects of the administered drug and simultaneously enhance its =
antitumor=20
      effect. There are 2 main concepts in DDS, active targeting and =
passive=20
      targeting. Active targeting involves a monoclonal antibody and a =
ligand to=20
      a tumor-related receptor. Doxil, a doxorubicine incorporated =
polyethylene=20
      glycol conjugated liposome, is categorized under passive targeting =
agents=20
      and are already in clinical use.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB13">13</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB14">14</A>]=20
      NK012, a novel SN-38-incorporating polymeric micelle, is a prodrug =
of=20
      SN-38 similar to CPT-11 and categorized under passive targeting =
agent as=20
      well. Although CPT-11 is converted to SN-38 in tumors by =
carboxylesterase=20
      (CE), the metabolic conversion rate is within 2-8% of the original =
volume=20
      of CPT-11.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB15">15</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB16">16</A>]=20
      In contrast, the release rate of SN-38 from NK012 is 74% under =
physiologic=20
      pH condition even without CEs.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB17">17</A>]=20
      Recently, we have demonstrated that NK012 exerted significantly =
more=20
      potent antitumor activity against various human tumor xenografts =
than=20
      CPT-11.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB17">17-20</A>]=20
      However, there is a fundamental question whether such =
nanoparticles can=20
      reach brain tumors across the tumor microvessels. In the present =
study,=20
      therefore, we established an orthotopic glioma model in this =
experiment=20
      and then evaluated whether NK012 can pass through the BTB and =
exert its=20
      antitumor effect on orthotopic human glioma xenografts in =
comparison with=20
      CPT-11.</P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC1-1>Material and =
methods</A></DIV>
      <TABLE>
        <TBODY>
        <TR>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#abstract"><IMG=20
            border=3D0 alt=3DAbstract=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><IMG border=3D0 alt=3D"Material and methods"=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_here_small.gif"></TD=
>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-2"><IMG=20
            border=3D0 alt=3DResults=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-3"><IMG=20
            border=3D0 alt=3DDiscussion=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#acknowledgements"><IMG=20
            border=3D0 alt=3DAcknowledgements=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#references"><IMG=20
            border=3D0 alt=3DReferences=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD></TR></TBODY></TABLE>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-1>Drugs</A></DIV>
      <P>NK012 and SN-38 were donated by Nippon Kayaku Co., Ltd. (Tokyo, =
Japan).=20
      The size of NK012 was <IMG border=3D0=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/sim.gif">20 nm =
in=20
      diameter with a narrow size distribution.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB17">17</A>]=20
      ACNU [1-(4-amino-2-methyl-5- pyrimidinyl)=20
      methyl-3-(2-chloroethyl)-3-nitrosourea, nimstine] was purchased =
from=20
      DAIICHI SANKYO Co., Ltd. (Tokyo, Japan). CDDP=20
      (cis-diamminedichloroplatinum) and CPT-11 were purchased from =
Yakult Co.,=20
      Ltd. (Tokyo, Japan). Etoposide [4<IMG border=3D0=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/prime.gif">demeth=
lepipodophyllotoxin-9-(4,=20
      6-O-ethylidene-<IMG border=3D0 align=3DabsBottom=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/beta.gif">-D-gluc=
opyranoside)]=20
      was purchased from BIOMOL (Plymouth Meeting, PA).</P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-2>Cells and =
animals</A></DIV>
      <P>Five human glioma cell lines, namely U87MG, U251MG, U118MG, =
LN18 and=20
      LN229, were obtained from the American Type Culture Collection =
(Rockville,=20
      MD). Cells were maintained in Dulbecco's modified Eagle's minimum=20
      essential medium supplemented with 10% fetal bovine serum (Cell =
Culture=20
      Technologies, Gaggenau-Hoerden, Germany), penicillin, streptomycin =
and=20
      amphotericin B (100 units/ml, 100 <IMG border=3D0 =
align=3DabsBottom=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">g/ml and =
25=20
      <IMG border=3D0 align=3DabsBottom=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">g/ml,=20
      respectively; Sigma, St. Louis, MO) in a humidified atmosphere =
containing=20
      5% CO<SUB>2</SUB> at 37=B0C. Six- to eight-week-old athymic nude =
mice=20
      (nu/nu; Charles River Japan, Kanagawa, Japan) were used for this =
study.=20
      U87MG cells (1 =D7 10<SUP>5</SUP>) were injected into the cerebral =

      hemisphere using a Hamilton syringe through an entry point 1 mm =
anterior=20
      and 1.8 mm lateral to the bregma to an intraparenchymal depth of =
2.5 mm.=20
      The rate of injection was 0.5 <IMG border=3D0 align=3DabsBottom=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l/min, =
and=20
      the needle was left in place for 5 min after completion of the =
injection.=20
      All animal procedures were performed in compliance with the =
Guidelines for=20
      the Care and Use of Experimental Animals established by the =
Committee for=20
      Animal Experimentation of the National Cancer Center, Japan; these =

      guidelines meet the ethical standards required by law and also =
comply with=20
      the guidelines for the use of experimental animals in Japan.</P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-3>Establishment of =
U87MG cell=20
      line stably expressing firefly luciferase and YFP mutant =
Venus</A></DIV>
      <P>For the <I>in vivo</I> bioluminescence imaging of orthotopic =
brain=20
      tumors, the U87MG cell line stably expressing firefly luciferase =
and the=20
      YFP mutant Venus was established. Briefly, the coding sequence for =
firefly=20
      luciferase and Venus was subcloned into the the pIRES Vector =
(Clontech=20
      Laboratories, Mountain View, CA). The fragment consists of=20
      Luciferase-IRES-Venus generated from the plasmid with the =
restriction=20
      enzymes Nhe I and Not I. This fragment was subcloned into the =
pEF6/V5-His=20
      Vector (Invitrogen, Carlsbad, CA) to generate plasmids of =
pEF6-Lucifease=20
      IRES Venus. U87MG cells (2 =D7 10<SUP>6</SUP>) were seeded onto =
10-cm dishes=20
      24 hr before transfection. The cells were transfected with 10 <IMG =

      border=3D0 align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">g =
of=20
      pEF6-Lucifease IRES Venus using FuGENE HD Transfection Reagent =
(Roche=20
      Diagnostics, Mannheim, Germany) according to manufacturer's =
instructions,=20
      and then incubated for 48 hr at 37=B0C. The cells were then =
passaged in=20
      medium containing Blasticidin (10 <IMG border=3D0 =
align=3DabsBottom=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">g/ml;=20
      InvivoGen, San Diego, CA) to select for the Blasticidin resistance =
gene=20
      integrated in the pEF6/V5-His plasmids. Venus expression was used =
as a=20
      surrogate marker of luciferase-positive cells. Venus-expressing =
U87MG=20
      cells (U87MG/Luc) were sorted using the BD FACS Aria cell sorter =
(BD=20
      Biosciences, San Jose, CA), and expanded in selection medium. The =
accuracy=20
      of a quantitive bioluminescence image as an indicator of U87MG/Luc =
cell=20
      number was analyzed using the Photon Imager animal imaging system =
<I>in=20
      vitro</I>, as described under <I>in vivo</I> growth inhibition =
assay. This=20
      analysis demonstrated clear correlation between a quantitive=20
      bioluminescence image and cell number (<I>R</I><SUP>2</SUP> =3D =
0.99). The=20
      sensitivity of U87MG/Luc cells to each drug (NK012, CPT-11, SN-38, =
ACNU,=20
      CDDP and etoposide) was almost similar to that of parental U87MG =
cells=20
      (data not shown).</P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-4>In vitro growth =
inhibition=20
      assay</A></DIV>
      <P>Cell growth inhibition was measured by the tetrazolium =
salt-based=20
      proliferation assay (WST assay; Wako Chemicals, Osaka, Japan). =
Briefly,=20
      cells (5 =D7 10<SUP>3</SUP> cells/well) in 96-well plates were =
incubated=20
      overnight. Then, growth medium was changed to new medium with =
various=20
      concentrations of SN-38, NK012, CPT-11, ACNU, etoposide and CDDP. =
After 72=20
      hr of incubation, medium was changed to new medium containing 10% =
WST-8=20
      reagents. After 1 hr of incubation, the absorbance of the formazan =
product=20
      formed was detected at 450 nm in a 96-well spectrophotometric =
plate reader=20
      (SpectraMax 190; Molecular Devices, Sunnyvale, CA). Cell viability =
was=20
      measured and compared with that of the control cells. Each =
experiment was=20
      carried out in triplicates and was repeated at least 3 times. Data =
were=20
      averaged and normalized against the nontreated controls to =
generate=20
      dose-response curves. The number of living cells (% Control) was=20
      calculated using the following formula: % Control =3D (each=20
      absorbance&nbsp;-&nbsp;absorbance of blank well)/absorbance of =
control=20
      well =D7 100.</P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-5>Evaluation of =
NK012 and CPT-11=20
      distribution in tumor tissue by fluorescence microscopy</A></DIV>
      <P>The U87MG orthotopic xenograft model described earlier was used =
for the=20
      analysis of the biodistribution of NK012 and CPT-11. Twenty days =
after=20
      U87MG/Luc inoculation, the maximum tolerated dose (MTD) of NK012 =
(30=20
      mg/kg) or CPT-11 (66.7 mg/kg) was injected intravenously into the =
tail=20
      vein of mice. At this point, tumor size reached to about 3 mm in =
diameter=20
      according to the preliminary experiment (data not shown). Two, 12 =
or 24 hr=20
      after NK012 or CPT-11 injection, mice were also administered with=20
      fluorescein <I>Lycopersicon esculentum</I> lectin (100 <IMG =
border=3D0=20
      align=3DabsBottom=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l/mouse) =

      (Vector Laboratories, Burlingame, CA) to visualize tumor blood =
vessels.=20
      Tumors were then excised and embedded in optimal cutting =
temperature=20
      compound and frozen at -80=B0C until use. Tissue sections (6 <IMG =
border=3D0=20
      align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">m =
thick)=20
      were prepared using Tissue-Tek Cryo3 (Sakura Finetek USA, Inc., =
Torrance,=20
      CA), and frozen sections were examined under a fluorescence =
microscope,=20
      BIOREVO BZ9000 (Keyence, Osaka, Japan), at an excitation =
wavelength of 377=20
      nm and an emission wavelength 447 nm to evaluate the distribution =
of=20
      CPT-11 and NK012 within the tumor tissues. Because formulations =
containing=20
      SN-38 bound <I>via</I> ester bonds possess a particular =
fluorescence, both=20
      CPT-11 and NK012 were detected under the same fluorescence =
conditions.=20
      Image data were recorded using BZ-II Analyzer 1.10 software =
(Keyence,=20
      Osaka, Japan).</P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-6>Pharmacokinetics =
study of=20
      NK012 and CPT-11</A></DIV>
      <P>Female BALB/c nude mice bearing U87MG/Luc tumor (<I>n</I> =3D =
3) were=20
      used for the analysis of the biodistribution of NK012 and CPT-11. =
Twenty=20
      days after the intracranial injection of U87MG/Luc cells, NK012 =
(30 mg/kg)=20
      or CPT-11 (66.7 mg/kg) was intravenously administered to the mice. =
Under=20
      anesthesia, blood, normal brain tissues and tumor tissues were =
obtained 2,=20
      12, 24 and 72 hr after NK012 or CPT-11 administration. Blood =
samples were=20
      collected in microtubes and immediately centrifuged at =
1,600<I>g</I> for=20
      15 min at 4=B0C. All samples were stored at -80=B0C until use.</P>
      <P>The normal brain and tumor samples were rinsed with physiologic =
0.9%=20
      NaCl solution, mixed with 0.1 M glycine-HCl buffer (pH =
3.0)/methanol at 5=20
      w/w%, and then homogenized. To analyze the concentration of free =
SN-38 and=20
      CPT-11, 100 <IMG border=3D0 align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of the=20
      tumor homogenates was mixed with 20 <IMG border=3D0 =
align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of 1 mM=20
      phosphoric acid/methanol (1:1), 40 <IMG border=3D0 =
align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of=20
      ultrapure water and 60 <IMG border=3D0 align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of=20
      camptothecin solution (10 ng/ml for SN-38 and 15 ng/ml for CPT-11) =
as an=20
      internal standard. To quantify free SN-38 and CPT-11 in plasma, 25 =
<IMG=20
      border=3D0 align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of plasma=20
      was mixed with 25 <IMG border=3D0 align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of 0.1 M=20
      HCl, and then added with 20 <IMG border=3D0 align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of 1 mM=20
      phosphoric acid/methanol (1:1) and 100 <IMG border=3D0 =
align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of CPT=20
      solution (10 ng/ml both for SN-38 and CPT-11). The samples were =
vortexed=20
      vigorously for 10 sec, and then filtered through Ultrafree-MC =
centrifugal=20
      filter devices with a cut-off molecular diameter of 0.45 <IMG =
border=3D0=20
      align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">m =
(Millipore=20
      Co., Bedford, MA). Reversed-phase HPLC was performed at 35=B0C on =
a=20
      Mightysil RP-18 GP column 150 =D7 4.6 mm<SUP>2</SUP> (Kanto =
Chemical Co.,=20
      Inc., Tokyo, Japan). Fifty microliters of a sample was injected =
into an=20
      Alliance Waters 2795 HPLC system (Waters, Milford, MA) equipped =
with a=20
      Waters 2475 multi <IMG border=3D0=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/lambda.gif">=20
      fluorescence detector. Fluorescence originating from SN-38 was =
detected at=20
      540 nm with an excitation wavelength of 365 nm and that =
originating from=20
      CPT-11 was detected at 430 nm with an excitation wavelength of 365 =
nm. The=20
      mobile phase was a mixture of 100 nmol/l ammonium acetate (pH 4.2) =
and=20
      methanol (11:9 (v/v)). The flow rate was 1.0 ml/min. The content =
of SN-38=20
      was calculated by measuring the relevant peak area and calibrating =
against=20
      the corresponding peak area derived from the CPT internal =
standard. Peak=20
      data were recorded using a chromatography management system =
(MassLynx=20
      v4.0, Waters).</P>
      <P>For polymer-bound SN-38 detection, SN-38 was released from the=20
      conjugate. Briefly, 20 <IMG border=3D0 align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of plasma=20
      and 100 <IMG border=3D0 align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of tissue=20
      samples were diluted with 20 <IMG border=3D0 align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of=20
      methanol (50%, v/v) and 20 <IMG border=3D0 align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of NaOH=20
      (0.3 mol/l for plasma and 0.7 mol/l for tissue). The samples were=20
      incubated for 15 min at 25=B0C. After incubation, 20 <IMG =
border=3D0=20
      align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of HCl=20
      (0.3 mol/l for plasma and 0.7 mol/l for tissue) and 60 <IMG =
border=3D0=20
      align=3DabsBottom=20
      src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">l =
of CPT=20
      solution (10 ng/ml for SN-38 and 15 ng/ml for CPT-11) were added =
to the=20
      samples, and then the hydrolysate was filtered through a =
MultiScreen=20
      Solvinert. Fifty microliters of the filtrate was applied to the =
same HPLC=20
      system as described earlier.</P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-7>In vivo growth =
inhibition=20
      assay</A></DIV>
      <P>
      <DIV class=3DsecondLevelHeading><A name=3DSEC3-1>Experiment =
1</A></DIV>
      <P>Six-week-old mice were subcutaneously inoculated with 1 =D7=20
      10<SUP>7</SUP> U87MG/Luc cells in the flank region. When tumor =
volume=20
      reached <IMG border=3D0=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/sim.gif">605=20
      mm<SUP>3</SUP>, mice were randomly divided into test groups =
consisting of=20
      3 mice per group (day 0). Drug was intravenously administered on =
days 0, 4=20
      and 8 into the tail vein. NK012 was administered at its MTD of 30=20
      mg/kg/day. The reference drug CPT-11 was given at its MTD of 66.7=20
      mg/kg/day in the optimal schedule reported.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB17">17</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB21">21</A>]=20
      The length (<I>i</I>) and width (<I>ii</I>) of tumor masses were =
measured=20
      twice a week, and tumor volume (TV) was calculated as follows: TV =
=3D (a =D7=20
      b<SUP>2</SUP>)/2. Relative tumor volumes (RTVs) at day <I>n</I> =
were=20
      calculated according to the following formula: RTV =3D =
TV<SUB><I>n</I></SUB>=20
      / TV<SUB>0</SUB>, where TV<SUB><I>n</I></SUB> is the tumor volume =
at day=20
      <I>n</I>, and TV<SUB>0</SUB> is the tumor volume at day 0.</P>
      <P>
      <DIV class=3DsecondLevelHeading><A name=3DSEC3-2>Experiment =
2</A></DIV>
      <P>To assess the antitumor effect of NK012 and CPT-11, <I>in =
vivo</I>=20
      bioluminescence imaging studies were performed using the Photon =
Imager=20
      animal imaging system (Biospace, Paris, France). For imaging, mice =
with=20
      intracranial U87MG/Luc tumor were simultaneously anesthetized with =

      isoflurane and <FONT size=3D-1>D</FONT>-luciferine potassium salt =
(Synchem,=20
      Germany), and normal 0.9% NaCl solution was intraperitoneally=20
      administrated at a dose of 125 mg/kg body weight, and images were =
obtained=20
      5 min after the injection. For bioluminescence image analysis, =
regions of=20
      interest encompassing the intracranial area of a signal were =
defined using=20
      Photo Vision software (Biospace, Paris, France), and total numbers =
of=20
      photons per minute (cpm) were recorded. The pseudo-color =
luminescent image=20
      from violet (least intense) to red (most intense) represented the =
spatial=20
      distribution of detected photon counts emerging from active =
luciferase=20
      within the animal. Twenty days after U87MG inoculation, treatment =
was=20
      started (day 0). Normal 0.9% NaCl solution (<I>n</I> =3D 4), NK012 =
(30=20
      mg/kg, <I>n</I> =3D 4), or CPT-11 (66.7 mg/kg, <I>n</I> =3D 4) was =

      intravenously administered to mice on days 0, 4 and 8. <I>In =
vivo</I>=20
      bioluminescence imaging studies were performed on days 0, 14, 21 =
and 28=20
      from the day of treatment initiation. To determine the effect of =
treatment=20
      on the time to change of intensity, Student's <I>t</I> test was =
carried=20
      out using the StatView 5.0 software package. <I>p</I> &lt; 0.05 =
was=20
      regarded as significant.</P>
      <P>
      <DIV class=3DsecondLevelHeading><A name=3DSEC3-3>Experiment =
3</A></DIV>
      <P>Mice with intracranial U87MG/Luc tumor was randomly divided =
into 3=20
      groups consisting of 6 mice per group. NK012 (30 mg/kg/day) and =
CPT-11=20
      (66.7 mg/kg/day) were intravenously given on days 0 (20 days after =
tumor=20
      inoculation), 4 and 8. After treatment, mice were maintained until =
each=20
      animal showed signs of morbidity (<I>i.e</I>., 10% weight loss and =

      neurological deficit), at which point they were sacrificed. =
Kaplan-Meier=20
      analysis was performed to determine the effect of drugs on time to =

      morbidity, and statistical differences were ranked according to =
the=20
      Mantel-Cox log-rank test using StatView 5.0.</P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-8>Statistical =
analysis</A></DIV>
      <P>Data were expressed as mean =B1 SD. Significance of differences =
was=20
      calculated using the unpaired <I>t</I> test with repeated measures =
of=20
      StatView 5.0. <I>p</I> &lt; 0.05 was regarded as significant.</P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC1-2>Results</A></DIV>
      <TABLE>
        <TBODY>
        <TR>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#abstract"><IMG=20
            border=3D0 alt=3DAbstract=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-1"><IMG=20
            border=3D0 alt=3D"Material and methods"=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><IMG border=3D0 alt=3DResults=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_here_small.gif"></TD=
>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-3"><IMG=20
            border=3D0 alt=3DDiscussion=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#acknowledgements"><IMG=20
            border=3D0 alt=3DAcknowledgements=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#references"><IMG=20
            border=3D0 alt=3DReferences=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD></TR></TBODY></TABLE>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-9>Cellular =
sensitivity of=20
      glioblastoma cells to SN-38, NK012 and CPT-11</A></DIV>
      <P>The IC<SUB>50</SUB> values of NK012 for the cell lines ranged =
from=20
      0.0087 <IMG border=3D0 align=3DabsBottom=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">mol/l=20
      (U251MG cells) to 0.36 <IMG border=3D0 align=3DabsBottom=20
      =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">mol/l =
(LN229=20
      cells). The growth inhibitory effects of NK012 were 34- to =
444-fold more=20
      potent than those of CPT-11, 400- to 12818-fold more potent than =
those of=20
      ACNU, 52-to 523-fold more potent than those of CDDP, and 3- to =
278-fold=20
      more potent than those of etoposide. On the other hand, the=20
      IC<SUB>50</SUB> values of NK012 were almost similar to those of =
SN-38=20
      (Table. <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#TBL1">I</A>).=20

      <CENTER>
      <TABLE width=3D"80%">
        <TBODY>
        <TR>
          <TD><A name=3DTBL1>&nbsp;</A></TD></TR>
        <TR>
          <TD align=3Dmiddle><B>Table I. </B><B>IC<SUB>50</SUB> Values =
of SN-38,=20
            NK012, CPT-11, ACNU, CDDP and VP-16 in Various Human =
Glioblastoma=20
            Cell Lines</B></TD></TR>
        <TR vAlign=3Dbottom>
          <TD>
            <HR SIZE=3D2 noShade>
          </TD></TR>
        <TR>
          <TD align=3Dmiddle>
            <TABLE>
              <TBODY>
              <TR>
                <TH vAlign=3Dbottom rowSpan=3D2 align=3Dleft>Cell =
line</TH>
                <TH vAlign=3Dbottom colSpan=3D6=20
                  align=3Dmiddle><NOBR>IC<SUB>50</SUB>(<IMG border=3D0=20
                  align=3DabsBottom=20
                  =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">mol/l)</N=
OBR>
                  <HR SIZE=3D1>
                </TH></TR>
              <TR>
                <TH vAlign=3Dbottom =
align=3Dmiddle><NOBR>SN38</NOBR></TH>
                <TH vAlign=3Dbottom =
align=3Dmiddle><NOBR>NK012</NOBR></TH>
                <TH vAlign=3Dbottom =
align=3Dmiddle><NOBR>CPT-11</NOBR></TH>
                <TH vAlign=3Dbottom =
align=3Dmiddle><NOBR>ACNU</NOBR></TH>
                <TH vAlign=3Dbottom =
align=3Dmiddle><NOBR>CDDP</NOBR></TH>
                <TH vAlign=3Dbottom =
align=3Dmiddle><NOBR>Etoposide</NOBR></TH></TR>
              <TR>
                <TD colSpan=3D7>
                  <HR SIZE=3D1>
                </TD></TR>
              <TR>
                <TD vAlign=3Dtop align=3Dleft><NOBR>LN18</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.052 =B1 0.0034</TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.069 =B1 0.0242</TD>
                <TD vAlign=3Dtop align=3Dmiddle>13.0 =B1 0.88</TD>
                <TD vAlign=3Dtop align=3Dmiddle>729 =B1 30</TD>
                <TD vAlign=3Dtop align=3Dmiddle>3.57 =B1 0.08</TD>
                <TD vAlign=3Dtop align=3Dmiddle>3.84 =B1 0.14</TD></TR>
              <TR>
                <TD vAlign=3Dtop align=3Dleft><NOBR>LN229</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.28 =B1 0.1094</TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.36 =B1 0.0489</TD>
                <TD vAlign=3Dtop align=3Dmiddle>12.2 =B1 1.23</TD>
                <TD vAlign=3Dtop align=3Dmiddle>144 =B1 23</TD>
                <TD vAlign=3Dtop align=3Dmiddle>21.4 =B1 0.62</TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.945 =B1 =
0.025</TD></TR>
              <TR>
                <TD vAlign=3Dtop align=3Dleft><NOBR>U87MG</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.18 =B1 0.0216</TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.093 =B1 0.0038</TD>
                <TD vAlign=3Dtop align=3Dmiddle>18.1 =B1 3.06</TD>
                <TD vAlign=3Dtop align=3Dmiddle>865 =B1 86</TD>
                <TD vAlign=3Dtop align=3Dmiddle>9.06 =B1 0.57</TD>
                <TD vAlign=3Dtop align=3Dmiddle>20.8 =B1 5.23</TD></TR>
              <TR>
                <TD vAlign=3Dtop align=3Dleft><NOBR>U118MG</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.0089 =B1 0.0003</TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.022 =B1 0.0017</TD>
                <TD vAlign=3Dtop align=3Dmiddle>4.85 =B1 0.14</TD>
                <TD vAlign=3Dtop align=3Dmiddle>282 =B1 22</TD>
                <TD vAlign=3Dtop align=3Dmiddle>3.35 =B1 0.35</TD>
                <TD vAlign=3Dtop align=3Dmiddle>4.05 =B1 0.18</TD></TR>
              <TR>
                <TD vAlign=3Dtop align=3Dleft><NOBR>U251MG</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.0076 =B1 0.0001</TD>
                <TD vAlign=3Dtop align=3Dmiddle>0.0087 =B1 0.0002</TD>
                <TD vAlign=3Dtop align=3Dmiddle>3.86 =B1 0.04</TD>
                <TD vAlign=3Dtop align=3Dmiddle>51.6 =B1 3.7</TD>
                <TD vAlign=3Dtop align=3Dmiddle>4.55 =B1 0.03</TD>
                <TD vAlign=3Dtop align=3Dmiddle>2.42 =B1=20
        0.13</TD></TR></TBODY></TABLE></TD></TR>
        <TR vAlign=3Dbottom>
          <TD>
            <HR SIZE=3D2 noShade>
          </TD></TR></TBODY></TABLE></CENTER>
      <CENTER>
      <TABLE width=3D"80%">
        <TBODY>
        <TR>
          <TD><FONT size=3D-1>&nbsp;&nbsp;&nbsp;Each cell line was =
treated in=20
            triplicate for 72 hr.<BR></FONT><FONT=20
            size=3D-1>&nbsp;&nbsp;&nbsp;WST-8 assay was used for =
obtaining=20
            IC<SUB>50</SUB> =
value.<BR></FONT></TD></TR></TBODY></TABLE></CENTER>
      <P></P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-10>Antitumor =
activity of NK012=20
      and CPT-11 on subcutaneous U87MG/Luc xenografts</A></DIV>
      <P>Potent antitumor activity was observed in mice treated with =
NK012 at 30=20
      mg/kg <I>in vivo</I> (Fig. <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#FIG1">1</A><I>a</I>).=20
      In mice treated with NK012, tumor volume started to decrease on =
day 5, and=20
      the tumor completely disappeared by day 23, with no relapse =
observed until=20
      80 days after treatment. Although CPT-11 at 66.7 mg/kg/day exerted =

      antitumor activity compared with the control group, tumor volume =
continued=20
      to increase consistently. Comparison of the relative tumor volume =
at day 8=20
      revealed significant differences between the NK012-treated and=20
      CPT-11-treated groups (<I>p</I> =3D 0.0095). Although =
treatment-related body=20
      weight loss was observed in mice treated with each drug, body =
weight=20
      recovery was observed by day 19 (Fig. <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#FIG1">1</A><I>b</I>).=20
      These results clearly show the significant <I>in vivo</I> activity =
of=20
      NK012 against the U87MG/Luc tumor xeonograft.=20
      <P><A name=3DFIG1></A></P>
      <CENTER>
      <TABLE border=3D0 width=3D"85%">
        <TBODY>
        <TR>
          <TD vAlign=3Dtop width=3D128 align=3Dleft><A=20
            href=3D"javascript:display_fig001(0)"><IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/tfig=
001"></A></TD>
          <TD align=3Dleft><FONT size=3D-1><B>Figure 1. </B>Effects of =
NK012 and=20
            CPT-11 on U87MG/Luc tumor xenograft. (<I>a</I>) Tumor volume =
in mice=20
            treated with CPT-11 or NK012. U87MG/Luc tumor was =
subcutaneously=20
            inoculated into the flank of mice, as described in the <A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-1">Materials=20
            and methods</A> section. NaCl (0.9%) solution (<IMG =
border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/bull.gif">),=20
            CPT-11 at 66.7 mg/kg (<IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/squf.gif">)=20
            and NK012 at 30 mg/kg (<IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/utrif.gif">)=20
            were intravenously administered on days 0, 4 and 8 (arrows). =
Points,=20
            mean; bars, SD. *<I>p</I> &lt; 0.05. (<I>b</I>) =
Treatment-related=20
            body weight loss occurred in mice treated with CPT-11 and =
NK012.=20
            Points, mean; bars, SD.<BR>[<A=20
            href=3D"javascript:display_fig001(0)">Normal View 22K</A> | =
<A=20
            href=3D"javascript:display_fig001(1)">Magnified View=20
        59K</A>]</FONT></TD></TR></TBODY></TABLE></CENTER>
      <P></P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-11>Studies on =
distribution of=20
      NK012 and CPT-11 in orthotopic U87MG/Luc tumor tissues</A></DIV>
      <P>Both NK012 and CPT-11 formulations accumulated in the tumor =
tissue but=20
      not in the normal brain tissue (Fig. <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#FIG2">2</A>).=20
      However, the drug distribution pattern was clearly different =
between NK012=20
      and CPT-11. In sections of the U87MG/Luc tumor treated with =
CPT-11,=20
      maximum drug accumulation was observed within 2 hr of CPT-11 =
injection.=20
      Twelve hours after the injection, fluorescence originating from =
CPT-11 had=20
      almost disappeared. Subsequently, no accumulation of CPT-11 was =
observed=20
      within the tumor tissues. However, in sections of the U87MG/Luc =
tumor=20
      treated with NK012, fluorescence from NK012 started appearing =
around tumor=20
      blood vessels 2 hr after intravenous injection and lasted until 24 =
hr.=20
      After 12 hr, the fluorescent area began to increase and the =
maximum=20
      fluorescence area was observed 24 hr after the injection.=20
      <P><A name=3DFIG2></A></P>
      <CENTER>
      <TABLE border=3D0 width=3D"85%">
        <TBODY>
        <TR>
          <TD vAlign=3Dtop width=3D128 align=3Dleft><A=20
            href=3D"javascript:display_fig002(0)"><IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/tfig=
002"></A></TD>
          <TD align=3Dleft><FONT size=3D-1><B>Figure 2. </B>Distribution =
of NK012=20
            or CPT-11 in U87MG/Luc glioma xenografts. Mice bearing =
U87MG/Luc=20
            tumor were injected with NK012 (30 mg/kg/day) or CPT-11 =
(66.7=20
            mg/kg/day). Tumor tissues were excised 2, 12 and 24 hr after =
the=20
            intravenous injection of NK012 or CPT-11. Each mouse was=20
            administered fluorescein-labeled <I>Lycopersicon =
esculentum</I>=20
            lectin 5 min before sacrifice to detect tumor blood vessels. =
Frozen=20
            sections were examined under a fluorescence microscope at an =

            excitation wavelength of 377 nm and an emission wavelength =
of 477=20
            nm. The same fluorescence conditions can be applied for =
visualizing=20
            NK012 and CPT-11 fluorescence. Free SN-38 could not be =
detected=20
            under these fluorescence conditions. The white lines =
indicate the=20
            border between the tumor and the brain tissue. T, U87MG/Luc =
tumor;=20
            B, normal brain tissue. (Scale bars: 20 <IMG border=3D0=20
            align=3DabsBottom=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/mu.gif">m).<BR>[<=
A=20
            href=3D"javascript:display_fig002(0)">Normal View 52K</A> | =
<A=20
            href=3D"javascript:display_fig002(1)">Magnified View=20
          283K</A>]</FONT></TD></TR></TBODY></TABLE></CENTER>
      <P></P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-12>Pharmacokinetics =
analysis of=20
      NK012 and CPT-11 in mice bearing orthotopic U87MG/Luc =
xenografts</A></DIV>
      <P>Microscopic observations were confirmed quantitatively by =
measuring the=20
      amount of SN-38 extracted from each solid tumor by reversed-phase =
HPLC.=20
      After CPT-11 injection, the concentrations of CPT-11 and free =
SN-38 in=20
      plasma decreased rapidly with time in a log-linear fashion. On the =
other=20
      hand, the plasma concentration of NK012 (polymer-bound SN-38) =
showed=20
      slower clearance than that of CPT-11. Free SN-38 released from =
NK012 also=20
      showed slow clearance than that of SN-38 converted from CPT-11 =
(Fig. <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#FIG3">3</A><I>a</I>).=20
      Meanwhile, there was a significant difference in drug accumulation =
in the=20
      tumor between CPT-11 and NK012, that is, the accumulation of NK012 =
in the=20
      U87MG/Luc tumor was significantly higher than that of CPT-11 (Fig. =
<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#FIG3">3</A><I>b</I>)=20
      and that the concentration of free SN-38 originating from NK012 =
was=20
      maintained at 24.6 ng/g even 72 hr after injection (Table <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#TBL2">II</A>).=20
      On the other hand, only slight conversion from CPT-11 to SN-38 was =

      observed from 2 to 24 hr in the U87MG/Luc tumor, and no SN-38 was =
detected=20
      thereafter (Fig. <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#FIG3">3</A><I>b</I>).=20
      This result suggests that the BTB of the tumor was partially =
destroyed in=20
      the tumor vasculature and both drugs extravasated from the tumor =
blood=20
      vessels. In addition, these results indicate that NK012 can remain =
in the=20
      tumor tissue for a longer period and continue to release free =
SN-38.=20
      <P><A name=3DFIG3></A></P>
      <CENTER>
      <TABLE border=3D0 width=3D"85%">
        <TBODY>
        <TR>
          <TD vAlign=3Dtop width=3D128 align=3Dleft><A=20
            href=3D"javascript:display_fig003(0)"><IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/tfig=
003"></A></TD>
          <TD align=3Dleft><FONT size=3D-1><B>Figure 3. </B>Plasma, =
brain tissue=20
            and orthotopic tumor concentrations of respective analytes =
after=20
            intravenous administration of NK012 (30 mg/kg/day) and =
CPT-11 (66.7=20
            mg/kg/day) to U87MG/Luc-bearing nude mice. (<I>a</I>) =
plasma;=20
            (<I>b</I>) tumor. <IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/bull.gif">,=20
            polymer-bound SN-38; <IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/cir.gif">,=20
            free SN-38 (polymer-unbound SN-38); <IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/utrif.gif">,=20
            CPT-11; <IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/Delta.gif">,=20
            free SN-38 converted from CPT-11.<BR>[<A=20
            href=3D"javascript:display_fig003(0)">Normal View 22K</A> | =
<A=20
            href=3D"javascript:display_fig003(1)">Magnified View=20
        59K</A>]</FONT></TD></TR></TBODY></TABLE></CENTER>
      <CENTER>
      <TABLE width=3D"80%">
        <TBODY>
        <TR>
          <TD><A name=3DTBL2>&nbsp;</A></TD></TR>
        <TR>
          <TD align=3Dmiddle><B>Table II. </B><B>Tumor and Plasma =
Concentration=20
            of SN38 After an I.V. Administration of NK012 (30 mg/kg) and =
CPT-11=20
            (66.7mg/kg) to Nude Mice Bearing U87MG/Luc Brain =
Tumor</B></TD></TR>
        <TR vAlign=3Dbottom>
          <TD>
            <HR SIZE=3D2 noShade>
          </TD></TR>
        <TR>
          <TD align=3Dmiddle>
            <TABLE>
              <TBODY>
              <TR>
                <TH vAlign=3Dbottom rowSpan=3D2 align=3Dleft>Formulation =
tested</TH>
                <TH vAlign=3Dbottom rowSpan=3D2=20
                align=3Dmiddle><NOBR>Analyte</NOBR></TH>
                <TH vAlign=3Dbottom rowSpan=3D2 =
align=3Dmiddle><NOBR></NOBR></TH>
                <TH vAlign=3Dbottom colSpan=3D4 align=3Dmiddle>Time =
after=20
                  administration (hr)
                  <HR SIZE=3D1>
                </TH></TR>
              <TR>
                <TH vAlign=3Dbottom align=3Dmiddle><NOBR>2</NOBR></TH>
                <TH vAlign=3Dbottom align=3Dmiddle><NOBR>12</NOBR></TH>
                <TH vAlign=3Dbottom align=3Dmiddle><NOBR>24</NOBR></TH>
                <TH vAlign=3Dbottom =
align=3Dmiddle><NOBR>72</NOBR></TH></TR>
              <TR>
                <TD colSpan=3D7>
                  <HR SIZE=3D1>
                </TD></TR>
              <TR>
                <TD vAlign=3Dtop rowSpan=3D2 =
align=3Dleft><NOBR>NK012</NOBR></TD>
                <TD vAlign=3Dtop rowSpan=3D2 align=3Dmiddle>Free =
SN-38</TD>
                <TD vAlign=3Dtop align=3Dleft>Plasma (ng/ml)</TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>1113</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>511</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>90.0</NOBR></TD>
                <TD vAlign=3Dtop =
align=3Dmiddle><NOBR>6.88</NOBR></TD></TR>
              <TR>
                <TD vAlign=3Dtop align=3Dleft>Tumor (ng/g)</TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>67.7</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>84.1</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>137</NOBR></TD>
                <TD vAlign=3Dtop =
align=3Dmiddle><NOBR>24.6</NOBR></TD></TR>
              <TR>
                <TD vAlign=3Dtop rowSpan=3D2 =
align=3Dleft><NOBR>CPT-11</NOBR></TD>
                <TD vAlign=3Dtop rowSpan=3D2 align=3Dmiddle>Free SN-38 =
<B>5</B></TD>
                <TD vAlign=3Dtop align=3Dleft>Plasma (ng/ml)</TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>62.0</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>4.74</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>1.97</NOBR></TD>
                <TD vAlign=3Dtop =
align=3Dmiddle><NOBR>ND</NOBR></TD></TR>
              <TR>
                <TD vAlign=3Dtop align=3Dleft>Tumor (ng/g)</TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>31.8</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>7.41</NOBR></TD>
                <TD vAlign=3Dtop align=3Dmiddle><NOBR>2.14</NOBR></TD>
                <TD vAlign=3Dtop=20
          =
align=3Dmiddle><NOBR>ND</NOBR></TD></TR></TBODY></TABLE></TD></TR>
        <TR vAlign=3Dbottom>
          <TD>
            <HR SIZE=3D2 noShade>
          </TD></TR></TBODY></TABLE></CENTER>
      <CENTER>
      <TABLE width=3D"80%">
        <TBODY>
        <TR>
          <TD><FONT size=3D-1>&nbsp;&nbsp;&nbsp;Data were expressed as =
means of=20
            three mice.<BR></FONT><FONT size=3D-1>&nbsp;&nbsp;&nbsp;Free =
SN-38;=20
            SN-38 released from NK012 or converted from =
CPT-11.<BR></FONT><FONT=20
            size=3D-1>&nbsp;&nbsp;&nbsp;ND, not=20
        detectable.<BR></FONT></TD></TR></TBODY></TABLE></CENTER>
      <P></P>
      <P>
      <DIV class=3DfirstLevelHeading><A name=3DSEC2-13>Antitumor =
activity of NK012=20
      and CPT-11 against orthotopic U87MG/Luc glioma =
xenografts</A></DIV>
      <P>Antitumor activity was observed in mice treated with NK012 at =
30=20
      mg/kg/day and CPT-11 at 66.7 mg/kg/day <I>in vivo</I> (Fig. <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#FIG4">4</A><I>a</I>).=20
      ANOVA analysis revealed a significant difference between the =
control group=20
      and the NK012-treated group (<I>p</I> =3D 0.02). However there was =
no=20
      significant difference between the control group and the =
CPT-11-treated=20
      group (<I>p</I> =3D 0.23) and between NK012 and CPT-11 (<I>p</I> =
=3D 0.21)=20
      (Fig. <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#FIG4">4</A><I>b</I>).=20
      Comparison of the relative tumor volume at day 14 revealed =
significant=20
      differences between NK012 (30 mg/kg/day) and CPT-11 (66.7 =
mg/kg/day)=20
      (<I>p</I> =3D 0.049). Kaplan-Meier analysis showed that a =
significant=20
      improvement in survival rate was observed in the NK012 treatment =
group=20
      compared with the control (<I>p</I> =3D 0.001) and CPT-11 =
treatment groups=20
      (<I>p</I> =3D 0.0014) (Fig. <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#FIG4">4</A><I>c</I>).=20

      <P><A name=3DFIG4></A></P>
      <CENTER>
      <TABLE border=3D0 width=3D"85%">
        <TBODY>
        <TR>
          <TD vAlign=3Dtop width=3D128 align=3Dleft><A=20
            href=3D"javascript:display_fig004(0)"><IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/tfig=
004"></A></TD>
          <TD align=3Dleft><FONT size=3D-1><B>Figure 4. </B>Antitumor =
effect of=20
            NK012 or CPT-11 on orthotopic xenograft and survival. Mice =
receiving=20
            intracranial injections of U87MG/Luc were assigned into =
groups 20=20
            days after tumor inoculation. Mice were intravenously =
administered=20
            with 0.9% NaCl solution (<IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/bull.gif">),=20
            NK012 (30 mg/kg/day, <IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/utrif.gif">)=20
            or CPT-11 (66.7 mg/kg/day, <IMG border=3D0=20
            =
src=3D"http://www3.interscience.wiley.com/giflibrary/12/squf.gif">) on=20
            days 0 (20 days after tumor inoculation), 4 and 8 (arrows).=20
            (<I>a</I>) Representative luminescence intensity images =
obtained in=20
            individual control and treatment-group mice on the days =
indicated.=20
            (<I>b</I>) Antitumor effect of NK012 or CPT-11 on days 14, =
21 and=20
            28. Each group consisted of 4 mice. Points, mean; bars, SD.=20
            *<I>p</I> &lt; 0.05. (<I>c</I>) Treatment effects of NK012 =
on=20
            survival. Survival was assessed by Kaplan-Meier analysis. =
Each group=20
            consisted of 6 mice. Experiments were repeated twice with =
similar=20
            results.<BR>[<A href=3D"javascript:display_fig004(0)">Normal =
View=20
            39K</A> | <A href=3D"javascript:display_fig004(1)">Magnified =
View=20
            114K</A>]</FONT></TD></TR></TBODY></TABLE></CENTER>
      <P></P>
      <P>
      <DIV class=3DfirstLevelHeading><A =
name=3DSEC1-3>Discussion</A></DIV>
      <TABLE>
        <TBODY>
        <TR>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#abstract"><IMG=20
            border=3D0 alt=3DAbstract=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-1"><IMG=20
            border=3D0 alt=3D"Material and methods"=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-2"><IMG=20
            border=3D0 alt=3DResults=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><IMG border=3D0 alt=3DDiscussion=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_here_small.gif"></TD=
>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#acknowledgements"><IMG=20
            border=3D0 alt=3DAcknowledgements=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#references"><IMG=20
            border=3D0 alt=3DReferences=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD></TR></TBODY></TABLE>
      <P>The diameter of a micelle carrier is approximately in the range =
of=20
      10-100 nm, which is smaller than that of a liposome. Although this =
size is=20
      small, it is still sufficiently large to prevent renal secretion =
of the=20
      carrier. The micelle systems can evade nonspecific capture by the=20
      reticuloendothelial system in various organs because the outer =
shell of=20
      the micelle is covered with polyethyleneglycol. Therefore,=20
      drug-incorporating micelles can be expected to have a long plasma=20
      half-life, which permits a large amount of the micelles to reach =
tumor=20
      tissues, extravasate from tumor capillaries, and then be retained =
in tumor=20
      tissues for a long time by utilizing the enhanced permeability and =

      retention (EPR) effect.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB22">22</A>]</P>
      <P>Several factors are reportedly involved in vascular =
permeability in the=20
      body. Among them, bradykinin is the most potent vascular =
permeability=20
      factor. We succeeded in purifying 2 types of kinin from the =
ascitic fluid=20
      of a patient with gastric cancer.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB23">23</A>]=20
      We also clarified that this kinin generation system was triggered =
by the=20
      activated Hageman factor, an intrinsic coagulation factor XII.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB24">24</A>]=20
      Meanwhile, Dvorak et al. discovered that the vascular permeability =
factor=20
      (VPF) is involved in tumor vascular permeability.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB25">25</A>]=20
      Later, it was found that VPF was identical to VEGF.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB26">26</A>]=20
      Recently, an extrinsic coagulation factor, namely, a tissue =
factor, has=20
      been shown to activate VEGF production.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB27">27</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB28">28</A>]=20
      Thus, both intrinsic and extrinsic coagulation factors may be =
involved in=20
      tumor vascular permeability. Furthermore, there have been several =
reports=20
      to date indicating the increasing expression of a tissue factor in =
human=20
      glioma.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB29">29</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB30">30</A>]=20
      Also, it is well known that glioma is a typical hypervascular =
tumor with=20
      an irregular vascular architecture and a high expression level of =
VEGF.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB31">31</A>]=20
      Therefore, it may be speculated that nanoparticles extravasate =
from tumor=20
      capillaries and accumulate more preferentially in brain =
glioma.</P>
      <P>NK012, an SN-38-incorporating polymeric micelle, is a novel =
type of=20
      micellar formulation with long-time accumulation in tumors, and =
shows=20
      prolonged sustained release of SN-38 within the tumor.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB17">17</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB20">20</A>]=20
      We have thus far reported that NK012 shows significantly higher =
antitumor=20
      activity against various human tumor xenografts including small =
cell lung=20
      cancer,[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB17">17</A>]=20
      colorectal cancer,[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB19">19</A>]=20
      renal cancer,[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB18">18</A>]=20
      and pancreatic cancer[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB20">20</A>]=20
      compared with CPT-11. In addition, we have recently seen an =
increasing=20
      number of reports of clinical trials indicating the effectiveness =
of=20
      CPT-11 against brain glioma in combination with anti-VEGF =
antibody[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB11">11</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB12">12</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB32">32</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB33">33</A>]</P>
      <P>Under these circumstances, it may be reasonable to conduct an=20
      investigation into the advantages of administering NK012 over =
CPT-11 for=20
      treatment against human glioma tumor xenografts. In the present =
study, we=20
      showed that NK012 exerted a significant antitumor activity in =
U87MG/Luc=20
      subcutaneous xenografts (Fig. <A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#FIG1">1</A>).=20
      In the tumor intravenously administered with NK012 (30 mg/kg), =
NK012=20
      accumulated within and around tumor blood vessels in the =
orthotopic=20
      xenografts 2 hr after the injection. Thereafter, NK012 started to =
spread=20
      from the blood vessel within the tumor tissue of the xenografts.=20
      Fluorescence originating from NK012 then increased up to the =
maximum in=20
      the entire tumor by 24 hr after NK012 injection. On the other =
hand,=20
      fluorescence originating from CPT-11 increased up to the maximum 2 =
hr=20
      after its injection, indicating that the maximum distribution of =
CPT-11=20
      was achieved within 2 hr of injection. Twelve hours after =
intravenous=20
      injection, fluorescence from CPT-11 had almost disappeared, and=20
      subsequently, no accumulation of CPT-11 was observed within the =
tumor=20
      tissues. The therapeutic effect of NK012 was superior to that of =
CPT-11 in=20
      terms of antitumor effect and survival. Because the antitumor =
activity of=20
      SN-38 is time-dependent, the superiority of NK012 over CPT-11 may =
be due=20
      to the enhanced accumulation of NK012 and the prolonged sustained =
release=20
      of SN-38 from NK012 within the tumor tissues. Nevertheless, free =
SN-38 was=20
      not detected in the normal brain tissues at any measurement time =
after=20
      intravenous injection of NK012 or CPT-11 (data not shown). It is =
thus=20
      speculated that both NK012 and CPT-11 are unable to cross the BBB =
in the=20
      normal brain, but can pass through tumor vessels effectively. In =
clinical=20
      brain glioma, however, when the tumor recurs, it would most likely =
recur=20
      in adjacent regions of the brain with an intact blood brain =
barrier.=20
      Namely, at the border between the brain tumor and normal brain =
tissue,=20
      malignant glioma cells and normal brain tissues intermix in the =
gradient=20
      and angiogenesis occurs at sites where there are large =
accumulation of=20
      tumor cells under local hypoxia.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB34">34</A>]=20
      In addition, there is no clear evidence if tumor vessels of =
orthotopic=20
      brain tumor xenografts are identical to those of real human brain =
tumors=20
      in terms of their structure and function. Therefore, it may be =
better to=20
      consider to conduct an investigation of the advantages of =
administering=20
      some anti-angiogenic inhibitors in combination with NK012 against =
highly=20
      invasive tumor models established by several methods such as =
direct=20
      implantation of patient surgical specimens into the brains of nude =

      mice,[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB35">35</A>]=20
      transplantation of patient surgical material s.c. in nude mice =
followed by=20
      dissociation and orthotopic reinjection of these =
xenotransplants,[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB36">36</A>]=20
      engraftment of glioblastoma-derived spheroids after short- term =
culture=20
      into rat brain,[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB37">37</A>]=20
      and engraftment of glioblastomastem cell-enriched cultures into =
mouse=20
      brain.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB38">38</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB39">39</A>]</P>
      <P>The dose-limiting toxicities of CPT-11 appeared to be =
neutropenia and=20
      diarrhea. However, in our previous data, there was no significant=20
      difference in the level of SN-38 in the small intestine between=20
      NK012-treated and CPT-11-treated mice.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB17">17</A>]=20
      It was also reported that NK012 showed significant antitumor =
effect with=20
      diminishing incidence of diarrhea compared with CPT-11.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB40">40</A>]=20
      In 2 individual phase 1 trials in Japan and the US, no serious =
diarrhea=20
      has been reported.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB41">41</A>][<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB42">42</A>]=20
      In addition, one confirmed partial response (PR) was obtained in a =
patient=20
      with metastatic esophageal cancer in a Japanese trial,[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB41">41</A>]=20
      and 3 PRs in a patient with breast cancer and 1 PR in a patient =
with small=20
      cell lung cancer in a US phase 1 trial.[<A=20
      =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#BIB42">42</A>]</P>
      <P>In conclusion, we demonstrated not only the enhanced =
accumulation,=20
      distribution, and retention of NK012 within glioma xenografts but =
also the=20
      superiority of the antitumor activity of NK012 compared with =
CPT-11.=20
      Taking the present data together with very recent clinical data =
from phase=20
      1 trials, a phase 2 trial in patients with recurrent glioma may be =

      warranted.</P>
      <P>
      <DIV class=3DfirstLevelHeading><A=20
      name=3Dacknowledgements>Acknowledgements</A></DIV>
      <TABLE>
        <TBODY>
        <TR>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#abstract"><IMG=20
            border=3D0 alt=3DAbstract=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-1"><IMG=20
            border=3D0 alt=3D"Material and methods"=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-2"><IMG=20
            border=3D0 alt=3DResults=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-3"><IMG=20
            border=3D0 alt=3DDiscussion=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><IMG border=3D0 alt=3DAcknowledgements=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_here_small.gif"></TD=
>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#references"><IMG=20
            border=3D0 alt=3DReferences=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_down_small.gif"></A>=
</TD></TR></TBODY></TABLE>
      <P>The authors thank Ms. N. Mie, Ms. H. Miyatake and Ms. M. Ohtsu =
for=20
      their technical assistance and Ms. K. Shiina for her secretarial=20
      assistance.</P>
      <P>
      <TABLE border=3D0 width=3D"100%">
        <TBODY>
        <TR>
          <TD class=3DmainSectionHeader><A=20
        name=3Dreferences>References</A></TD></TR></TBODY></TABLE>
      <TABLE>
        <TBODY>
        <TR>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#abstract"><IMG=20
            border=3D0 alt=3DAbstract=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-1"><IMG=20
            border=3D0 alt=3D"Material and methods"=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-2"><IMG=20
            border=3D0 alt=3DResults=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#SEC1-3"><IMG=20
            border=3D0 alt=3DDiscussion=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><A=20
            =
href=3D"http://www3.interscience.wiley.com/cgi-bin/fulltext/121534411/mai=
n.html,ftx_abs#acknowledgements"><IMG=20
            border=3D0 alt=3DAcknowledgements=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_up_small.gif"></A></=
TD>
          <TD><IMG border=3D0 alt=3DReferences=20
            =
src=3D"http://www3.interscience.wiley.com/images/sec_here_small.gif"></TD=
></TR></TBODY></TABLE>
      <TABLE width=3D"100%">
        <TBODY>
        <TR vAlign=3Dbaseline>
          <TD class=3DreferenceNum>1</TD>
          <TD><A name=3DBIB1>DeAngelis LM. Brain tumors. <I>N Engl J =
Med</I>=20
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