Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/adodb/adodb.inc.php on line 893

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/adodb/adodb.inc.php on line 1918

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/adodb/adodb.inc.php on line 1990

Deprecated: mysql_connect(): The mysql extension is deprecated and will be removed in the future: use mysqli or PDO instead in /www/htdocs/w00a10d9/html/includes/classes/adodb/drivers/adodb-mysql.inc.php on line 360

Deprecated: Function split() is deprecated in /www/htdocs/w00a10d9/html/includes/pnLang.php on line 62

Strict Standards: Declaration of pnRender::clear_cache() should be compatible with Smarty::clear_cache($tpl_file = NULL, $cache_id = NULL, $compile_id = NULL, $exp_time = NULL) in /www/htdocs/w00a10d9/html/includes/pnRender.class.php on line 45

Strict Standards: Declaration of pnRender::is_cached() should be compatible with Smarty::is_cached($tpl_file, $cache_id = NULL, $compile_id = NULL) in /www/htdocs/w00a10d9/html/includes/pnRender.class.php on line 45

Strict Standards: Declaration of pnRender::fetch() should be compatible with Smarty::fetch($resource_name, $cache_id = NULL, $compile_id = NULL, $display = false) in /www/htdocs/w00a10d9/html/includes/pnRender.class.php on line 45

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 41

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 42

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 43

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 44

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 45

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 46

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 47

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 48

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 49

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 50

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 51

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 52

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 53

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 54

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 41

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 42

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 43

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 44

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 45

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 46

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 47

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 48

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 49

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 50

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 51

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 52

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 53

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 54

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 41

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 42

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 43

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 44

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 45

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 46

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 47

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 48

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 49

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 50

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 51

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 52

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 53

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 54

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 41

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 42

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 43

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 44

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 45

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 46

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 47

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 48

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 49

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 50

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 51

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 52

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 53

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 54

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 41

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 42

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 43

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 44

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 45

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 46

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 47

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 48

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 49

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 50

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 51

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 52

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 53

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/pnAntiCracker.php on line 54

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/safehtml.php on line 607

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 161

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 163

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 164

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 165

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 167

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 168

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 169

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 244

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 251

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 260

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 267

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 274

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 281

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 289

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 486

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 488

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/includes/classes/safehtml/HTMLSax3.php on line 490

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 51

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 51

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 51

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 51

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 51

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 53

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 54

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 55

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 56

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 57

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 58

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 63

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 64

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 64

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 65

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 66

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 67

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 68

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 69

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 70

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/includes/legacy/counter.php on line 71

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/modules/Xanthia/pnuserapi.php on line 70

Deprecated: Assigning the return value of new by reference is deprecated in /www/htdocs/w00a10d9/html/modules/Xanthia/pnuserapi.php on line 119

Strict Standards: Declaration of pnCEHTML::URL() should be compatible with pnHTML::URL($url = '', $text = '') in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 238

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 16

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 18

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 20

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 22

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 45

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 47

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 48

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 50

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 52

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 54

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 56

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 58

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 60

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 62

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 16

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 18

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 20

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 22

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 45

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 47

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 48

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 50

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 52

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 54

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 56

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 58

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 60

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 62

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/header.php on line 30
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Strict Standards: Non-static method ADOConnection::UnixTimeStamp() should not be called statically, assuming $this from incompatible context in /www/htdocs/w00a10d9/html/includes/classes/adodb/adodb.inc.php on line 3007

Strict Standards: Non-static method ADOConnection::UnixTimeStamp() should not be called statically, assuming $this from incompatible context in /www/htdocs/w00a10d9/html/includes/classes/adodb/adodb.inc.php on line 3007

Strict Standards: Non-static method ADOConnection::UnixTimeStamp() should not be called statically, assuming $this from incompatible context in /www/htdocs/w00a10d9/html/includes/classes/adodb/adodb.inc.php on line 3007

Strict Standards: Non-static method ADOConnection::UnixTimeStamp() should not be called statically, assuming $this from incompatible context in /www/htdocs/w00a10d9/html/includes/classes/adodb/adodb.inc.php on line 3007

Strict Standards: Non-static method ADOConnection::UnixTimeStamp() should not be called statically, assuming $this from incompatible context in /www/htdocs/w00a10d9/html/includes/classes/adodb/adodb.inc.php on line 3007

Strict Standards: Non-static method ADOConnection::UnixTimeStamp() should not be called statically, assuming $this from incompatible context in /www/htdocs/w00a10d9/html/includes/classes/adodb/adodb.inc.php on line 3007

Strict Standards: Non-static method ADOConnection::UnixTimeStamp() should not be called statically, assuming $this from incompatible context in /www/htdocs/w00a10d9/html/includes/classes/adodb/adodb.inc.php on line 3007

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 16

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 18

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 20

Deprecated: Function eregi() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 22

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 45

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 47

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 48

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 50

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 52

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 54

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 56

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 58

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 60

Deprecated: Function ereg() is deprecated in /www/htdocs/w00a10d9/html/modules/ContentExpress/pneditor/pnCEHTML.php on line 62

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

Deprecated: preg_replace(): The /e modifier is deprecated, use preg_replace_callback instead in /www/htdocs/w00a10d9/html/includes/pnAPI.php on line 589

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The Development of Immunology in the Austro-Hungarian Empire and in the First Republic

H. Eibl
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Soon after Behring had discovered how to use the blood or blood serum of immunized animals to transfer their immunity against a certain infection to other animals, and after the first patients had successfully been treated with immune sera obtained from horses, institutions for the production of such immune sera were founded in various European countries, and also in the Austro-Hungarian Empire, where, on the initiative of Paltauf, a Lyssa- and a Diphtheria Seruminstitute, the predecessor of the Serotherapeutic Institute were founded in Vienna.

In the purely academic field it was Max von Gruber who accepted an appointment as head of the new Institute of Hygiene in Vienna. Gruber became well-known for his discovery of the agglutination of typhoid and cholera bacilli by sera of infected animals and humans. The same discovery was independently made by Widal in 1897, which is why this diagnostic principle which is still applied today is called the Gruber-Widal-reaction.

Although Paltauf's own contributions were mainly in the fields of pathology and experimental pathology, several of his assistants such as Kraus and Pick, and later Eisler-Terramare, were most intensively involved in the field of Immunology. In 1897, Kraus discovered precipitation. After having passed bacterial lysates through a bacteria-proof filter he observed that filtrates and the corresponding antisera - regardless of whether they had been obtained from humans or immunized animals - formed precipitates. This showed that not only bacteria but also their soluble components react with antibodies of human or animal origin and that visible precipitates may form in the process.

Another very important antigen-antibody reaction was toxin-neutralization by antibodies. Ehrlich had experience from earlier studies with the plant toxin ricin, which enabled him to develop a theoretical basis for the neutralization of toxins by specific antibodies within the context of the side-chain theory he had postulated.

While Paltauf and his assistants agreed fully to this theory as the basis of toxin-antitoxin-neutralization, Gruber could not subscribe to this view and thus - intentionally or unintentionally - was in sharp contradiction to of Paul Ehrlich whose views were resolutely advocated by Paltauf and his disciples.

A young physician, Karl Landsteiner, who was then but 28 years old and had had excellent training in chemistry, successfully applied for a job at the newly established Institute of Hygiene under Max von Gruber. Landsteiner was born in Vienna in 1868. His father died seven years later. In 1885 he completed high school and commenced his university studies. In 1890, shortly before his graduation, he was baptized together with his mother in the Schottenkirche and named Karl Otto. A few months later, he received his medical degree. His broad range of interests took him to other very well known chemical laboratories at different universities. He worked as doctor-in-training at medical and surgical clinics.

Landsteiner, at a very early stage, developed his own views on scientific matters, which is why his opinion on the antigen-antibody-reaction differed distinctly from that of Paul Ehrlich and his school. Essentially, Landsteiner tended towards Gruber's point of view, but developed his own hypothesis which was shared by Max von Gruber even when Landsteiner left the Institute of Hygiene one year later in 1897. Landsteiner joined the Institute of Pathology as trainee-assistant, where, after a few months, he even got a salary, and less than a year later became assistant to the famous bacteriologist Weichselbaum who proposed him for habilitation in 1903. His work at the Institute of Pathology was extremely intensive. In addition to a series of important discoveries, he discoverd and succeeded in investigating the human blood groups and isoagglutinins in detail and thus established the human blood group system. In the ten years of his work he performed more than 3500 autopsies as well.

The dispute over Paul Ehrlich's side-chain theory as an explanation of the antigen-antibody reaction continued. Landsteiner repeatedly pointed out that this reaction and the chemical binding between antigen and antibody assumed by Paul Ehrlich and his co-workers, in particular by Morgenroth, did not take place. The different opinions led to debates between Max von Gruber and the group of Paul Ehrlich involving not only Ehrlich's co-workers but also Paltauf and his assistants. These conflicts had already become apparent when he gave his commemoration speech on the demise of Pasteur in 1895, because Gruber had always tended more towards the scientific way of thinking of Pasteur and his co-workers than towards that of the circle of researchers in Berlin and Frankfurt around Robert Koch and Paul Ehrlich; since the German-French war, considerable animosities had developed between these groups. As a consequence of the disputes, which had escalated in 1901 and 1902 Max von Gruber accepted an offer of the Institute of Hygiene in Munich and left Vienna.

One of Paltauf's young co-workers, Löwenstein, worked on a difficult problem. The bacterial cultures used to immunize animals caused very severe local and systemic reactions, which sometimes led to severe illnesses or death in these animals.
Even in his early investigations, Behring tried to mitigate the effect of bacterial toxins by treating them with various substances in order to make safe immunization of the animals possible, but without real success.

The detoxification of toxins by means of carbon disulphide claimed by Ehrlich turned out to be unreproducible, too.

At first, among the many disinfectants used at that time, none could be found which would mitigate the adverse reaction without destroying the antigenic effect. The first to succeed was Löwenstein who treated toxins with formol and intensive light of different colors and finally came to the conclusion that toxin treated with formaldehyde and subsequently irradiated with red light is detoxicated while retaining at least part of its antigenic effect.

At about the same time, after his discovery of the blood groups, Landsteiner in 1904 studied other haemagglutinin systems, some of them together with Eisler-Terramare. The latter, however subsequently devoted himself to a different field at the Serotherapeutic Institute. Due to his good training in physics and chemistry, he managed - together with Löwenstein - to prove that to detoxify bacterial, plant, and animal toxins - if they are proteins - by means of formaldehyde, it is not necessary to irradiate them with red light, but that the essential factor in this detoxification is heat. Landsteiner and Eisler-Terramare continued to work together in this field, and after several years succeeded in systematically developing formol toxoids, so that constant results of animal experiments were obtained and it was finally possible to plan the immunization of large animals and humans.

From 1908 onward, Landsteiner systematically developed his ideas on antigens and their reaction with antibodies, working together with the colloid chemist Wolfgang Pauli - the father of the physicist and Nobel laureate Pauli.

In co-operation with the cardiologist-to-be Jagitsch, Landsteiner first performed a series of basic studies on the reactions of colloidal substances and their possible relevance in antigen-antibody reaction. Later, together with Pauli, he succeeded in developing an electrophoresis apparatus, which showed the migration of antigens, antibodies and their complexes in the electric field. Essentially, this apparatus was the basis for the electrophoresis equipment developed a quarter of a century later bay the Nobel-prize winner Tiselius.

Landsteiner also repeatedly pointed out that antigen-antibody reactions were essentially reactions between colloids and that the chemical binding - which today we would call covalent binding - described by Ehrlich in his side-chain theory did not take place.

These different views repeatedly led to concealed and sometimes open disputes in publications and other scientific reports.

In 1908, Landsteiner left the Institute of Pathology. He accepted a position as prosector at the Wilheminenspital where he devoted his time to animal experiments for the intensive investigation of syphilis and poliomyelitis. After all other animal species had proved unsuitable, he managed to transfer syphilis to monkeys. In those animals not infected in these studies he tried to produce poliomyelitis by parenterally transferring autopsy material from patients who had died from poliomyelitis to these animals. A Cynomolgus monkey injected with the spinal cord of a child who had died from poliomyelitis showed paralyses and died after a few days; another experimental animal only showed paralyses. The significance of these experiments is in Landsteiner's findings that the histologic changes characteristic of poliomyelitis are present in the central nervous system of both animals. Together with Erwin Popper, an assistant at the Department of Pediatrics at the Wilhelminenspital, Landsteiner presented these results to the Medical Society in 1908. They were published in the following year.

Landsteiner continued his animal experiments in the field of poliomyelitis with Levaditi from the Institute Pasteur. Based on these studies about 50 years later a safe vaccine against poliomyelitis could be produced. Production of the Salk vaccine was stopped following the disastrous series of infections in 1955 and 1956 but taken up again later after the problem of complete inactivation was solved.

Two other contemporary scientists contributed to the field of immunology: Paltauf's assistant Pick and the pediatrician Pirquet who had returned from the John-Hopkins-University in Baltimore where he had held a chair in pediatrics. At the beginning of his work, Pick was still under the influence of Ehrlich's side-chain theory, but then dislinked himself increasingly from Ehrlich's ideas and tended more and more towards Landsteiner's point of view. Subsequently, by means of chemical modification, he succeeded in detecting new antigenic specificities. In 1906 he left the circle around Paltauf and became assistant at the Institute of Pharmacology where he continued his immunological studies.

Pirquet developed his ideas based on the observation that repeated treatment with substances of antigenic character may led to various different local and systemic reactions. For this phenomenon, he introduced the term "allergy". With "Pirquet's reaction" - named after him - he succeeded, with the help of tuberculin, to demonstrate a past or protracted active infection with tubercle bacilli in patients.

The Austro-Hungarian Empire was about to enter into World War I which, understandably, would result in an extensive restructuring of medical research. As prosector and head of pathology of the first Military Hospital, Landsteiner was overworked with autopsies. Eisler-Terramare and Löwenstein tried for the first time to apply their formol toxoid in humans. The results of animal experiments obtained by Löwenstein enabled Eisler-Terramare to vaccinate a small group of soldiers in the Austro-Hungarian army with tetanus formol toxoid.

At first, the results were not satisfactory, since the antitoxin titers reached in guinea pigs could not be achieved in man. In the immunization of horses and small animals they were used to reach titers of 100 and more units per milliliter serum. Today we know that this trial was indeed very successful, but that at that time they did not know that one hundred of an antitoxin unit per milliliter serum is enough to successfully protect a person against tetanus.

The end of World War I, especially the last two years of this war, left terrible marks. Hirszfeld, who visited Vienna in 1919, described Landsteiner as a human wreck on account of his chronic malnutrition.

In 1919, Landsteiner was told that his salary from the Viennese Hospital Fund would be stopped as of December 31st, and that future payments should come from the province of Lower Austria. Landsteiner had married in 1915 and had a son, Karl-Ernst, born in 1917. He felt, according to Eisler-Terramare, he should turn his back on his home country, most of all because of the disastrous under-supply, which, due to the acute shortage of milk, affected especially small children. He decided to accept a position as prosector of the Catholic Hospital in Den Haag. On top of everything else, he was informed that on May 20th of the following year he would be retired - with an income which due to inflation was so low that he would not be able to cover even the barest necessities of life.

His time in Holland, during which, despite every hardship, he continued his scientific work and his publications, was rather ill-starred. He had to carry out his laboratory work in a room in which the hospital's urinalyses were performed and which also served as a coffee room for the staff. The only help occasionally at his disposal was a nun whose time was utterly limited on account of breaks for prayers and her activity as organist in the hospital chapel. Still, in addition to his work as prosector, Landsteiner was able to publish nine scientific papers between 1919 and 1921. Although the conditions of life in Holland were considerably better than in Austria, he accepted the invitation by Simon Flexner, the director of the Rockefeller Institute, to work at this important scientific institution.

After the end of the War, Eisler-Terramare continued his work under Paltauf at the Serotherapeutic Institute. Rudolf Kraus, who had left Austria shortly before the outbreak of World War I, spent ten years in South America, mostly in Argentina and Brazil and had performed important microbiological studies and trained a number of young scientists, returned to the Serotherapeutic Institute in 1923.

In 1924 Paltauf died, and the financial situation of the Serotherapeutic Institute - now state owned - was disastrous. Under these conditions, Eisler-Terramare was unable to develop the formol toxoids invented by him and Löwenstein further in order to produce veterinary and human vaccines. Instead, Ramon and his co-workers at the Institute Pasteur as well as Glenny and co-workers were the ones to continue this work and to be extremely successful in developing formol toxoids into vaccines suitable for human use after the end of World War I.

Ramon did no longer use the word "toxoids", a term from Ehrlich's side-chain theory, but called the formol toxoids obtained from bacterial toxins as anatoxins. He managed to develop a method for the exact quantitation of these anatoxins, purified them and obtained vaccines which could be applied in humans and in animals.

For various reasons, immune sera against diphtheria obtained from immunized horses were only used for therapy, but not for prophylaxis, while mixtures of diphtheria toxins with diphtheria antitoxins obtained from horses, which had been developed by Behring, were administered as vaccines against diphtheria even before World War I. The morbidity rate as well as mortality due to diphtheria was still very high, so the search for a suitable vaccine continued.

In 1925 there was a terrible accident in Baden. Small children who had been vaccinated with such diphtheria toxin - antitoxin vaccines died of diphtheria-toxin intoxication. Immediately, intensive efforts were undertaken to elucidate the incident. The head of government control, Busson, who was responsible for the release of these vaccines, claimed that the diphtheria toxin - antitoxin mixture used had disintegrated and released the toxin. However, Grassberger, the head of the Institute of Hygiene, who had been commissioned as expert, pointed out that no horse protein had been serologically detected in these incriminated vaccines, so that probably the source material had been mixed up in the course of manufacturing and no antitoxin added. There was a violent argument between Busson and Grassberger in the Viennese Medical Society, which ended in Grassberger leaving the Medical Society. The whole incident was never completely clarified.

Incidents after vaccination against diphtheria similar to those in Austria also occurred in the USA, and detailed examinations of such toxin-antitoxin mixtures proved that these complexes could be broken up by physical changes. Further studies showed that the incidents in the USA apparently had occurred because neutralized toxin-antitoxin mixtures had become toxic after they had been simply frozen and thawed.

In principle, there was always the potential danger of toxicity of such neutralized toxin-antitoxin mixtures. On the basis of Ehrlich's ideas, however, they were at first considered low-risk vaccines, since it was assumed that solid chemical bond between antigen and antibody existed.

This view was contradicted by Landsteiner and Pauli, and later also by Pick, who claimed that this was not a usual kind of chemical bond, but a reaction between colloids. Even several years before the incident in Baden, three years after the death of Paul Ehrlich in 1914, his former assistant Morgenroth slated that there was no longer any evidence that the antigen-antibody reaction was a chemical reaction in the usual sense. Thus, Landsteiner's opinion had finally gained acceptance.

In the years to come, the Serotherapeutic Institute was not able to purify diphtheria formol toxoid sufficiently to prevent the severe local and systemic reactions. Löwenstein attempted percutaneous immunization with an ointment containing diphtheria toxoid. However, the results of this attempt varied so greatly in the vaccines that the possibility of percutaneous immunization was not pursued any further.

In 1930, after having been granted the US citizenship in 1929, Landsteiner - to his own surprise - was awarded the Nobel prize for his discovery made 30 years before on the blood group characteristics of human erythrocytes and their corresponding isoagglutinins.

To his co-workers, Landsteiner commented that he would have been more pleased if he had received the Nobel prize for the systematic detection of determinant groups in antigens. He thought that this special field of studies which he had initiated during his time in Vienna and which had been pursued by Pick and his co-workers independently from him, was of greater scientific importance than the discovery of human blood group characteristics.

Landsteiner was not very happy with his work at the Rockefeller Institute. Speaking with his co-workers, he called Simon Flexner, the head of the Institute, an "emperor" who would not allow him to go beyond his field of work which in his opinion is rather narrow. He also considered the constant obligation to report on his own activities and that of his co-workers unnecessary. Flexner also prevented Landsteiner from continuing his work in the field of experimental poliomyelitis research; one may well speculate whether the polio vaccine disaster of 1955/56 could have been prevented if Landsteiner had had the chance to continue the work he had begun in Vienna at the Rockefeller Institute.

In 1939, Landsteiner was conferred emeritus status, but got the chance to continue his work at the Rockefeller Institute, where in 1943 he suffered a severe heart attack and two days later died at the age of 75 years.

His wish for sufficient independence as head of a scientific institute was never fulfilled, just as Eisler-Terramare, who, after the death of Kraus in 1932 succeeded him as scientific head of the Serotherapeutic Institute, never won the Nobel prize for the invention of formol toxoids.

Due to the economic depression, political turmoil and the emigration of researchers, immunological research in Central Europe was surpassed by the immunological research in the USA in the period between the Wars. A comparison of the number of publications in the "Zeitschrift für Immunitätsforschung" and in the "Journal of Immunology" clearly shows the increasing number of contributions from Anglo-American laboratories.

In the thirties, the state-owned Serotherapeutic Institute was leased to the Serum-Union, a company dominated by Hungarians, and at first was able to hold its ground exonomically. The "Anschluss" of Austria to the German Reich put an end to this development. The company and thus the Serotherapeutic Institute, too, were taken over by non jewish owners and finally became the property of the Farbwerke Hoechst/Behringwerke. The senior staff dispersed.

World War II had started. The soldiers of the Allied Forces, in contrast to the forces of the Axis Powers were immunized with tetanus formol toxoid vaccines. The production sites of the previous Serotherapeutic Institute still produced only horse serum. Eisler-Terramare was taken into a concentration camp and, miraculously, survived. After the end of World War II he became scientific head of the Bundesstaatliches Serotherapeutisches Institut established by the Second Republic and held this position for over 10 years. In the course of the development of the Second Republic, scientific and technological research in the field of immunology successfully raised to international level and thus Austria regained the position it had held before World War I.

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