Strict Standards: Non-static method SEBenchmark::getInstance() should not be called statically in /home3/nextron/public_html/header.php on line 35

Strict Standards: Non-static method SESmarty::getInstance() should not be called statically in /home3/nextron/public_html/header.php on line 43

Strict Standards: Non-static method SEDatabase::getInstance() should not be called statically in /home3/nextron/public_html/header.php on line 96

Strict Standards: Non-static method SELanguage::info() should not be called statically in /home3/nextron/public_html/header.php on line 104

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 334

Strict Standards: Non-static method SELanguageStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 156

Strict Standards: Declaration of SELanguageStorageMysql::edit() should be compatible with SELanguageStorage::edit() in /home3/nextron/public_html/include/language/storage/mysql.php on line 255

Strict Standards: Declaration of SELanguageStorageMysql::get() should be compatible with SELanguageStorage::get() in /home3/nextron/public_html/include/language/storage/mysql.php on line 255

Strict Standards: Non-static method SECore::getSettings() should not be called statically in /home3/nextron/public_html/header.php on line 107

Strict Standards: Non-static method SECache::getInstance() should not be called statically in /home3/nextron/public_html/include/class_core.php on line 28

Strict Standards: Non-static method SECacheStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/cache/cache.php on line 66

Strict Standards: Non-static method SEDatabase::getInstance() should not be called statically in /home3/nextron/public_html/include/class_core.php on line 39

Strict Standards: Non-static method SECache::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_url.php on line 67

Strict Standards: Non-static method SECacheStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/cache/cache.php on line 66

Strict Standards: Non-static method SEDatabase::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_url.php on line 78

Strict Standards: Non-static method SELanguage::_preload_multi() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_datetime.php on line 35

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 529

Strict Standards: Non-static method SESession::getInstance() should not be called statically in /home3/nextron/public_html/header.php on line 125

Strict Standards: Non-static method SESessionStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/session/session.php on line 49

Strict Standards: Declaration of SESessionStorageNone::register() should be compatible with SESessionStorage::register($options = Array) in /home3/nextron/public_html/include/session/storage/none.php on line 16

Strict Standards: Non-static method SEDatabase::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/session/session.php on line 600

Strict Standards: Non-static method SESession::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_user.php on line 509

Strict Standards: Non-static method SESession::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_admin.php on line 208

Strict Standards: Non-static method SELanguage::select() should not be called statically in /home3/nextron/public_html/header.php on line 181

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 234

Strict Standards: Non-static method SELanguage::info() should not be called statically in /home3/nextron/public_html/header.php on line 183

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 334

Strict Standards: Non-static method SELanguage::info() should not be called statically in /home3/nextron/public_html/header.php on line 189

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 334

Strict Standards: Non-static method SECore::getPlugins() should not be called statically in /home3/nextron/public_html/header.php on line 229

Strict Standards: Non-static method SECache::getInstance() should not be called statically in /home3/nextron/public_html/include/class_core.php on line 62

Strict Standards: Non-static method SECacheStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/cache/cache.php on line 66

Strict Standards: Non-static method SEDatabase::getInstance() should not be called statically in /home3/nextron/public_html/include/class_core.php on line 73

Strict Standards: Declaration of rc_rssfeed::delete() should be compatible with rc_model::delete($id = NULL) in /home3/nextron/public_html/include/class_rssfeed.php on line 132

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 720

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 1087

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 1595

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 1636

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 1656

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 1756

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 1960

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2358

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2362

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2366

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2370

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2418

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2440

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2445

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2449

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2453

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2501

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2523

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2927

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2935

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2943

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2951

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 2959

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3300

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3304

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3308

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3312

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3373

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3395

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3434

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3456

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3461

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3465

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3469

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3473

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3783

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3815

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3845

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3868

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3879

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3889

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3911

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3925

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3952

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 3978

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4167

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4180

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4201

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4214

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4242

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4256

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4278

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4292

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4496

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4532

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4568

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4594

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4623

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4637

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4668

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4698

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4851

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4876

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4908

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4934

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 4988

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5117

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5153

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5186

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5217

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5321

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5353

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5392

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5428

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5463

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5498

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5505

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5790

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5794

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5798

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5802

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5850

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5872

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5877

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5881

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5885

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5933

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 5955

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 6392

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 7588

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 7649

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 7730

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 12921

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 12986

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 13011

Deprecated: Assigning the return value of new by reference is deprecated in /home3/nextron/public_html/include/class_radcodes_rss.php on line 13574

Strict Standards: Non-static method SELanguage::_preload_multi() should not be called statically in /home3/nextron/public_html/header_rssfeed.php on line 10

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 529

Strict Standards: Non-static method rc_toolkit::get_setting() should not be called statically in /home3/nextron/public_html/header_rssfeed.php on line 14

Strict Standards: Non-static method rc_toolkit::owner_level() should not be called statically in /home3/nextron/public_html/header_rssfeed.php on line 25

Strict Standards: Non-static method SELanguage::_preload_multi() should not be called statically in /home3/nextron/public_html/header_article.php on line 10

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 529

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_article.php on line 66

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_article.php on line 67

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SELanguage::_preload() should not be called statically in /home3/nextron/public_html/header_event.php on line 15

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 559

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_event.php on line 67

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_event.php on line 69

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_event.php on line 71

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_event.php on line 73

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_event.php on line 75

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SELanguage::_preload() should not be called statically in /home3/nextron/public_html/header_group.php on line 14

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 559

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_group.php on line 82

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_group.php on line 85

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_group.php on line 88

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SELanguage::_preload() should not be called statically in /home3/nextron/public_html/header_classified.php on line 15

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 559

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_classified.php on line 64

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_classified.php on line 66

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_classified.php on line 68

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_radcodes.php on line 10

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SELanguage::_preload_multi() should not be called statically in /home3/nextron/public_html/header_vid.php on line 15

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 529

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_vid.php on line 44

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SE_Hook::register() should not be called statically in /home3/nextron/public_html/header_vid.php on line 45

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 151

Strict Standards: Non-static method SELanguage::_preload_multi() should not be called statically in /home3/nextron/public_html/header_page.php on line 11

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 529

Strict Standards: Non-static method SELanguage::_preload_multi() should not be called statically in /home3/nextron/public_html/header_gmap.php on line 10

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 529

Strict Standards: Non-static method SE_Hook::exists() should not be called statically in /home3/nextron/public_html/header.php on line 298

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 246

Strict Standards: Non-static method SECache::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_user.php on line 280

Strict Standards: Non-static method SECacheStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/cache/cache.php on line 66

Strict Standards: Non-static method SEDatabase::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_user.php on line 291

Strict Standards: Non-static method SECache::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_user.php on line 320

Strict Standards: Non-static method SECacheStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/cache/cache.php on line 66

Strict Standards: Non-static method SEDatabase::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_user.php on line 331

Strict Standards: Non-static method SECache::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_user.php on line 200

Strict Standards: Non-static method SECacheStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/cache/cache.php on line 66

Strict Standards: Non-static method SEDatabase::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_user.php on line 211

Strict Standards: Non-static method SECache::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_user.php on line 240

Strict Standards: Non-static method SECacheStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/cache/cache.php on line 66

Strict Standards: Non-static method SEDatabase::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_user.php on line 251

Strict Standards: Non-static method SELanguage::_preload() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/class_user.php on line 179

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 559

Strict Standards: Non-static method SECore::getLanguages() should not be called statically in /home3/nextron/public_html/footer.php on line 14

Strict Standards: Non-static method SECache::getInstance() should not be called statically in /home3/nextron/public_html/include/class_core.php on line 96

Strict Standards: Non-static method SECacheStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/cache/cache.php on line 66

Strict Standards: Non-static method SELanguage::_languages() should not be called statically in /home3/nextron/public_html/include/class_core.php on line 110

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 366

Strict Standards: Non-static method SE_Hook::exists() should not be called statically in /home3/nextron/public_html/footer.php on line 29

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 246

Strict Standards: Non-static method SE_Hook::exists() should not be called statically in /home3/nextron/public_html/footer.php on line 32

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 246

Strict Standards: Non-static method SE_Hook::call() should not be called statically in /home3/nextron/public_html/footer.php on line 32

Strict Standards: Non-static method SE_Hook::create() should not be called statically in /home3/nextron/public_html/include/class_hook.php on line 272

Strict Standards: Non-static method SELanguage::info() should not be called statically in /home3/nextron/public_html/footer.php on line 58

Strict Standards: Non-static method SELanguage::_init() should not be called statically in /home3/nextron/public_html/include/language/language.php on line 334

Strict Standards: Non-static method SELanguage::_preload_multi() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/smarty/templates_c/%%86^861^861EA998%%article.tpl.php on line 6

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 529

Strict Standards: Non-static method SELanguage::load() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/smarty/templates_c/%%86^861^861EA998%%article.tpl.php on line 7

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 686

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/storage.php on line 192

Strict Standards: Non-static method SECache::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/storage.php on line 193

Strict Standards: Non-static method SECacheStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/cache/cache.php on line 66

Strict Standards: Only variables should be passed by reference in /home3/nextron/public_html/include/language/storage/mysql.php on line 177

Strict Standards: Non-static method SELanguage::_preload_multi() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/smarty/templates_c/%%05^053^053AB562%%header_article2.tpl.php on line 4

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 529

Strict Standards: Non-static method SELanguage::load() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/smarty/templates_c/%%05^053^053AB562%%header_article2.tpl.php on line 5

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 686

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/storage.php on line 169

Strict Standards: Non-static method SECache::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/storage.php on line 170

Strict Standards: Non-static method SECacheStorage::getInstance() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/cache/cache.php on line 66
Researchers turn Salmonella into anti-viral gene therapy agent - <br /> <b>Strict Standards</b>: Non-static method SELanguage::_get() should not be called statically, assuming $this from incompatible context in <b>/home3/nextron/public_html/include/smarty/templates_c/%%05^053^053AB562%%header_article2.tpl.php</b> on line <b>16</b><br /> <br /> <b>Strict Standards</b>: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in <b>/home3/nextron/public_html/include/language/language.php</b> on line <b>614</b><br /> insciences
Strict Standards: Non-static method SELanguage::_preload_multi() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/smarty/templates_c/%%87^870^870F1A81%%headerx.tpl.php on line 6

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 529

Strict Standards: Non-static method SELanguage::load() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/smarty/templates_c/%%87^870^870F1A81%%headerx.tpl.php on line 7

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 686
Keyword Tag Sort by


Categories: Salmonella Anti-viral gene therapy Virology Bacteria Ribozymes RNase P ribozyme Cytomegalovirus CMV Viruses Gene Therapy

Researchers turn Salmonella into anti-viral gene therapy agent

BERKELEY — New experiments at the University of California, Berkeley, may one day lead to anti-viral treatments that involve swallowing Salmonella bacteria, effectively using one bug to stop another.

Researchers at UC Berkeley’s School of Public Health have reprogrammed Salmonella, the same foodborne pathogen that can cause diarrhea, fever and abdominal cramps, into a safe transport vehicle for virus-stopping enzymes. Not only did this technique effectively treat mice infected with cytomegalovirus, it worked as an oral solution that was swallowed instead of injected.

Virologist Fenyong Liu teamed up with bacteriologist Sangwei Lu to develop the innovative technique, which is described in a study to be published online the week of Feb. 7 in the journal Proceedings of the National Academy of Sciences.

“A number of vaccines, including those for polio and smallpox, use live but weakened viruses to build up the immune system. But this is the first time anyone has successfully engineered bacteria for treatment of a viral infection,” said Liu, a UC Berkeley professor at the Division of Infectious Diseases & Vaccinology.

The researchers said Salmonella was particularly appealing because it has evolved to survive the human digestive system, allowing it to be swallowed instead of injected or inhaled.

“This is the first gene therapy treatment for viral infection that can be taken by mouth, which is far more convenient to administer than an injection,” said Lu, a UC Berkeley associate adjunct professor at the Division of Infectious Diseases & Vaccinology. “Moreover, there is already an attenuated strain of Salmonella with a decent track record for safety in humans since it is now used in the vaccine for typhoid (a disease caused by Salmonella Typhi).”

Researchers know that ribozymes, enzymes that are able to target and cut specific RNA molecules, can be used to inactivate a pathogen’s genes. But to do their work, ribozymes need to first get into the cells, and for that they need help.

It so happens that Salmonella is very good at invading cells, so the researchers found a way to use the bacterium as a vector for the RNase P ribozyme that could stop the gene activity of cytomegalovirus, or CMV.

CMV is in the same family of herpes viruses that causes cold sores, mononucleosis and chickenpox. CMV infections are generally mild among healthy individuals, but they can become deadly for people whose immune systems are compromised and are a leading viral cause of mental retardation in newborns.

Previous research by Liu and Lu showed that Salmonella could effectively sneak the anti-viral ribozymes into human cells infected with human cytomegalovirus and reduce the viral load of the cell cultures. This new study put the technique to the test in living mice.

As an added measure of safety, researchers took the attenuated strain of Salmonella and further mutated a gene that the bacteria needs to replicate. They tested the new mutant Salmonella strain in mice and confirmed that the mice did not get sick.

They then cloned the anti-viral ribozymes into a plasmid, or DNA molecules within the bacteria that can replicate. Among mice that had been infected with cytomegalovirus, those that had been given oral doses of the ribozyme-carrying Salmonella survived much better than mice that had not been treated or mice that had been given Salmonella carrying a defective version of the ribozyme. The treated mice lived at least 50 days after infection, whereas the mice in the other two groups died within 25 days after infection.

Moreover, the researchers found that the viral load of mice treated with the ribozyme-carrying Salmonella was 400- to 600-times lower than the viral load for mice given the defective ribozymes and for mice that were untreated.

The researchers pointed out that using bacteria instead of viruses as gene-therapy vectors has a number of advantages.

“Viruses can’t replicate on their own; they must be grown in host cells,” said Lu. “It is more challenging to grow host cells in a lab, and there is always the risk that those cells can be contaminated with unknown viruses. To grow bacteria, you only need to add some bacteria to a simple medium, and the next day you can have 100 billion bacteria ready to go. It’s safer, easier and cheaper as a vector for gene therapy.”

The researchers pointed to the potential for developing this technique into a range of gene-targeting therapeutics. “This study focused on the use of Salmonella and ribozymes to fight infections, but with more research, this method could eventually be used to treat other conditions as well, including cancer,” said Liu.

Other UC Berkeley authors on the paper include lead author Yong Bai and Hao Gong, both post-doctoral researchers in infectious diseases; Hongjian Li, a former post-doctoral researcher in infectious diseases; and Gia-Phong Vu, a graduate student in comparative biochemistry.

The U.S. Department of Agriculture and the National Institutes of Health helped support this research.

Sarah Yang

Contact: Sarah Yang, Tel: (510) 643-7741, Email: scyang@berkeley.edu

Source: UC Berkeley ( Berkeley, University of California)

Photos (1)  [ view all photos ]


Shown is an image of Salmonella bacteria taken by an electron microscope. Researchers at UC Berkeley have turned this foodborne pathogen into an anti-viral delivery agent. (Sangwei Lu image)

Related News:

Detour leads to antibiotic resistance 1 April 2014, 05:54
LMU researchers have used cryo-electron microscopic imaging to characterize the structural...

Single mutation gives virus new target 22 October 2013, 03:41
A mutation as minute as swapping just one amino acid can completely change the target that a...

Bugs without borders: Researchers track the emergence and global... 10 December 2012, 08:34
Researchers show that the global epidemic of Clostridium difficile 027/NAP1/BI in the early to...

Scientists discover new method of gene identification 12 November 2012, 02:46
Findings will help our genetic understanding of dangerous new viruses.Scientists studying the...

Mozzie Protein Alert to Invading Viruses 2 October 2012, 04:02
CSIRO scientists have discovered how mosquitoes develop viral immunity, potentially leading to...

Researchers Halt Autoimmune Disease Myasthenia Gravis in Mice 1 October 2012, 13:44
Johns Hopkins team says genetic therapy uses targeted “guided missile” cells, erasing need...

Novel Pathogen Epidemic Identified in Sub-Saharan Africa 1 October 2012, 04:29
Researchers track the spread of human invasive non-Typhoidal Salmonella in sub-Saharan Africa.A...

New African cassava resists devastating viruses 26 September 2012, 03:41
Plant scientists at ETH Zurich have developed a new African cassava preferred by consumers and...

How immune cells defend themselves against HIV 26 September 2012, 03:36
Heidelberg virologists decode natural blockade mechanism in the target cells of the HIV...

New Study Confirms Erroneous Link Between XMRV and Prostate... 19 September 2012, 04:59
Study Confirms Erroneous Link Between Prostate Cancer and Retrovirus from Mice.A once-promising...

Credits / Source:

Shown is an image of Salmonella bacteria taken by an electron microscope. Researchers at UC Berkeley have turned this foodborne pathogen into an anti-viral delivery agent. (Sangwei Lu image)

Sign up for daily newsletter

Subscribe to daily Science News
(a free e-mail delivery powered by Google)

Top Categories



Strict Standards: Non-static method SELanguage::load() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/smarty/templates_c/%%47^479^479B3B2F%%insciences.tpl.php on line 4

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 686

image

We have launched our peer- reviewed, Open-access journal: Insciences Journal

Recently added

27.11.12
Consequential Life Cycle Assessment of Nitrogen Fertilisers Based on Biomass – a Swedish perspective

08.11.12
From Molecular Hollows to Imprinting Strategies – Mass-Sensitive Devices in Host-Guest Chemistry

26.04.12
CdSe/ZnSe Quantum-Dot Semiconductor Optical Amplifiers

23.03.12
Screen-printed Electrochemical Sensors and Biosensors for Monitoring Metal Pollutants


 
 

Strict Standards: Non-static method SELanguage::load() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/smarty/templates_c/%%76^762^76224BF7%%footerx.tpl.php on line 6

Strict Standards: Non-static method SELanguage::_init() should not be called statically, assuming $this from incompatible context in /home3/nextron/public_html/include/language/language.php on line 686