Studying the effect of single mismatches in primer and probe binding regions on amplification curves and quantification in real-time PCR.

Comparison of a broad range of characteristics of real-time PCR amplification curves yielded only slight alterations for low numbers of mismatches in the primer binding regions, resulting in a quantification error up to 63.12%. The effects were more pronounced for mismatches in the probe binding region and resulted in a quantification error up to 33%.

Metabolic engineering of omega3-very long chain polyunsaturated fatty acid production by an exclusively acyl-CoA-dependent pathway.

omega3-Very long chain polyunsaturated fatty acids (VLCPUFA) are essential for human development and brain function and, thus, are indispensable components of the human diet. The current main source of VLCPUFAs is represented by ocean fish stocks, which are in severe decline, and the development of alternative, sustainable sources of VLCPUFAs is urgently required. Our research aims at exploiting the powerful infrastructure available for the large scale culture of oilseed crops, such as rapeseed, to produce VLCPUFAs such as eicosapentaenoic acid in transgenic plants.

Monitoring hygiene on- and at-line is critical for controlling Listeria monocytogenes during produce processing.

The prevalence of Listeria monocytogenes in different types of produce and on processing plant environments was investigated over a 4-year period in a large produce processing plant in Poland. Prevalence of L. monocytogenes was 46% in frozen vegetables and 41.

Strongyloides stercoralis: a rare cause of obstructive duodenal stenosis.

Background: Strongyloidiasis is a rare helminthic infection in Europe, and it may cause duodenal obstruction.

Methods: We report a patient who was admitted to our Medical Department with nausea and repeated vomiting since 10 years until food intake became impossible. Subsequent investigations revealed a duodenal obstruction at the upper third of the duodenum, as well as enterocolitis of the terminal ileum with eosinophils dispersed throughout the mucosa.

Selective expansion of the beta-cell compartment in the pancreas of keratinocyte growth factor transgenic mice.

Epithelial-mesenchymal interactions are essential for growth, differentiation, and regeneration of exocrine and endocrine cells in the pancreas. The keratinocyte growth factor (KGF) is derived from mesenchyme and has been shown to promote epithelial cell differentiation and proliferation in a paracrine fashion. Here, we have examined the effect of ectopic expression of KGF on pancreatic differentiation and proliferation in transgenic mice by using the proximal elastase promoter.

Clinical hepatotoxicity. Regulation and treatment with inducers of transport and cofactors.

Drug-induced liver injury (DILI) is a rare condition. This review presents a clinical update on the incidence and prevalence of DILI, cellular targets and histopathological features of liver injury, and basic diagnostic approaches. At present, with the exception of acetaminophen poisoning where timely administered N-acetylcysteine serves as the antidote, treatment usually is supportive.

Ethidium monoazide and propidium monoazide for elimination of unspecific DNA background in quantitative universal real-time PCR.

Unspecific background DNA in quantitative universal real-time PCR utilizing a hydrolysis probe was completely suppressed by the addition of EMA or PMA to the PCR mix via cross-linking of the dyes to DNA during 650 W visible light exposure. The proposed procedure had no effect on the sensitivity of the real-time PCR reaction.

Pancreatic stellate cells potentiate proinvasive effects of SERPINE2 expression in pancreatic cancer xenograft tumors.

We have previously reported that inducible overexpression of the serine protease inhibitor nexin 2 (SERPINE2) significantly increases local invasiveness of subclones of the pancreatic cancer cell-line SUIT-2 in nude mouse xenografts. This was associated with a striking increase of extracellular matrix deposition in the invasive tumors. Pancreatic stellate cells (PSCs) have recently been identified as the major source of fibrosis in pancreatic adenocarcinomas.

Expression of bile acid synthesis and detoxification enzymes and the alternative bile acid efflux pump MRP4 in patients with primary biliary cirrhosis.

Background: Bile acid synthesis, transport and metabolism are markedly altered in experimental cholestasis. Whether such coordinated regulation exists in human cholestatic diseases is unclear. We therefore investigated expression of genes for bile acid synthesis, detoxification and alternative basolateral export and regulatory nuclear factors in primary biliary cirrhosis (PBC).

Fluorescence and ultraviolet absorption spectra, and the structure and vibrations of 1,2,3,4-tetrahydronaphthalene in its S1(pi,pi*) state.

The ultraviolet absorption spectra in the static vapor phase and the laser induced fluorescence spectra (both fluorescence excitation and single vibronic level fluorescence spectra) of jet-cooled 1,2,3,4-tetrahydronaphthalene have been used along with theoretical calculations to assign many of the vibronic levels in the S1(pi,pi*) state. These have been compared to the corresponding vibrational levels for the S0 ground state. Analysis of the upper states of the ring-twisting vibration nu(31) and three other low-frequency modes has allowed us to construct an energy map of the lowest vibrational quantum states for both S0 and S1.

Endocrine modulation and toxic effects of two commonly used UV screens on the aquatic invertebrates Potamopyrgus antipodarum and Lumbriculus variegatus.

The two UV screens 3-benzylidene-camphor (3-BC) and 3-(4'-methylbenzylidene)-camphor (4-MBC) were tested regarding their toxicity and estrogenic activity. The Yeast Estrogen Screen (YES) and two sediment assays with the freshwater invertebrates Lumbriculus variegatus and Potamopyrgus antipodarum were performed. In the YES, both substances activated the human estrogen receptor alpha with EC50 values of 44.

Atorvastatin in patients with primary biliary cirrhosis and incomplete biochemical response to ursodeoxycholic acid.

Unlabelled: Statin therapy may target both hypercholesterolemia and cholestasis in primary biliary cirrhosis (PBC). However, little is known about the efficacy and safety of statins in PBC. The aim of this single-center study was therefore to prospectively examine the effects of atorvastatin on serum markers of cholestasis, aminotransferases, and lipid and bile acid metabolism as well as inflammatory and immunological markers in patients with PBC.

Hepatobiliary transporter expression in intercellular adhesion molecule 1 knockout and Fas receptor-deficient mice after common bile duct ligation is independent of the degree of inflammation and oxidative stress.

Liver injury in intercellular adhesion molecule 1 knockout (ICAM(-/-)) and Fas receptor-deficient (lpr) mice is markedly reduced after common bile duct ligation (CBDL) due to significantly reduced inflammation and oxidative stress. Liver injury in CBDL rodents is counteracted by adaptive hepatobiliary transporter induction. Since hepatobiliary transporter expression in obstructive cholestasis may be regulated not only by accumulating bile acids but also by inflammatory mediators and oxidative stress, we hypothesized that differences in the inflammatory response may affect hepatobiliary transporter expression in CBDL, which would contribute to reduced liver injury.

Overexpression of ErbB2 in the exocrine pancreas induces an inflammatory response but not increased proliferation.

Tyrosine kinase receptors such as members of the epidermal growth factor receptor family and their respective ligands are frequently overexpressed in pancreatic cancer as well as in chronic pancreatitis. In this study, the role of ErbB2 in the exocrine pancreas was examined by ectopic overexpression under the control of the proximal rat elastase promoter. Three independent transgenic mouse lines overexpressing ErbB2 were established by pronuclear injection.

Constitutive IKK2 activation in acinar cells is sufficient to induce pancreatitis in vivo.

Activation of the inhibitor of NF-kappaB kinase/NF-kappaB (IKK/NF-kappaB) system and expression of proinflammatory mediators are major events in acute pancreatitis. However, the in vivo consequences of IKK activation on the onset and progression of acute pancreatitis remain unclear. Therefore, we modulated IKK activity conditionally in pancreatic acinar cells.

Development of matrix lysis for concentration of gram positive bacteria from food and blood.

The development of a fast, reliable and inexpensive protocol for the concentration of bacteria from food by the removal of fat, carbohydrates and proteins that is compatible with downstream alternative DNA-based quantification methods is described. The protocol was used for dairy products, cooked and smoked fish and meat, carbohydrate-rich cooked products, ready-to-eat sauces, egg and blood. Lysis resulted in pellets of reasonable size for further processing.

Real-time PCR method with statistical analysis to compare the potential of DNA isolation methods to remove PCR inhibitors from samples for diagnostic PCR.

A real-time PCR method for fast comparison of different DNA isolation methods to remove PCR inhibitors from samples is presented. A fixed amount of target-200 copies of a 79-bp region of the COCH gene of the zebrafish (Danio rerio)-was added to each PCR reaction together with isolated DNA from different types of samples including chicken feces. Four commercial DNA isolation kits and a chelex-based technique were compared using this method.

MDR3 (ABCB4) defects: a paradigm for the genetics of adult cholestatic syndromes.

Because ATP-binding cassette (ABC) transporters are important for normal bile secretion, hereditary and acquired ABC transporter defects play a central role in the pathogenesis of cholestasis. Defects of the phospholipid export pump MDR3 ( ABCC4) result in impaired biliary excretion of phosphatidylcholine and a variety of cholestatic syndromes ranging from progressive familial intrahepatic cholestasis in neonates to biliary cirrhosis in adults. Moreover, MDR3 mutations predispose to cholestasis of pregnancy and drug-induced cholestasis.

Principles of hepatic organic anion transporter regulation during cholestasis, inflammation and liver regeneration.

Hepatic uptake and biliary excretion of organic anions (e.g., bile acids and bilirubin) is mediated by hepatobiliary transport systems.

Analysis of apoptosome dysregulation in pancreatic cancer and of its role in chemoresistance.

The apoptosome is a multiprotein complex mediating the mitochondrial pathway of cell death. Its importance during development has been clearly demonstrated by knocking out key genes in mouse. APAF1 is the core protein of the apoptosome and its dosage is also critical in various cancer types, i.