Expression of epithelial cell adhesion molecule (EpCam) in renal epithelial tumors

EpCam is an epithelial adhesion molecule expressed in a broad range of carcinomas. Clinical trials with specific humanized anti-EpCam antibodies have shown promising results and have been inaugurated in renal cell carcinoma (RCC) therapy. To study the EpCam expression profile, primary renal cell neoplasms as well as corresponding metastases were evaluated by immunohistochemistry in tissue microarrays. EpCam expression in oncocytomas and chromophobe RCCs was determined on conventional large sections. Moderate or strong EpCam expression was found in eighteen percent of clear cell (n=147), 75% of chromophobe (n=12), and 55% of papillary RCCs (n=20), but not in oncocytomas (n=3). On large sections, 90% of chromophobe RCCs (n=20) showed a strong and homogeneous positivity, whereas oncocytomas (n=15) revealed EpCam positivity in single tumor cells or small clusters. Fourteen percent of RCC metastases (n=97) showed EpCam expression. Patients with EpCam expressing clear cell RCC showed a trend toward a better prognosis in a Cox regression analysis including stage, grade, and necrosis. The data suggest EpCam as a potential therapeutic target in a subset of patients with RCC. In addition, expression patterns of EpCam could become a helpful tool in the discrimination of chromophobe RCC and oncocytoma.     

Prenatal exposure to the CB1 receptor agonist WIN 55,212-2 causes learning disruption associated with impaired cortical NMDA receptor function and emotional reactivity changes in rat offspring

The aim of this study was to investigate whether prenatal exposure to the cannabinoid CB1 receptor agonist WIN 55,212-2 (WIN) at a daily dose devoid of overt signs of toxicity and/or gross malformations (0.5 mg/kg, gestation days 5-20), influences cortical glutamatergic neurotransmission, learning and emotional reactivity in rat offspring. Basal and K+-evoked extracellular glutamate levels were significantly lower in cortical cell cultures obtained from pups exposed to WIN during gestation with respect to those measured in cultures obtained from neonates born from vehicle-treated dams. The addition of NMDA to cortical cell cultures from neonates born from vehicle-treated dams concentration-dependently increased glutamate levels, and this was absent in cell cultures obtained from WIN-exposed pups. WIN-exposed rats also revealed a poorer performance in homing (10-12 days of age) and active avoidance tests (80 days of age) as well as a decrease in the rate of separation-induced ultrasonic emission (10 days of age). Finally, prenatal exposure to WIN induced a reduction in the number of cortical neuronal population. These findings (i) provide evidence for a deficit in cortical glutamatergic neurotransmission and behaviour in the rat neonate following prenatal exposure to WIN; and (ii) suggest that the reduction in cortical glutamatergic neurotransmission, NMDA receptor activity and alterations in neuronal development might underlie, at least in part, the learning deficit and decreased emotional reactivity observed in the offspring.     

Mitochondrial type I nitric oxide synthase physically interacts with cytochrome c oxidase

Nitric oxide (NO) regulates key aspects of cell metabolism through reversible inhibition of cytochrome c oxidase (CcOX), the terminal electron acceptor (complex IV) of the mitochondrial respiratory chain, in competition with oxygen. Recently, a constitutive mitochondrial NOS corresponding to a neuronal NOS-I isoform (mtNOS-I) has been identified in several tissues. The role of this enzyme might be to generate NO close enough to its target without a significant overall increase in cellular NO concentrations. An effective, selective, and specific NO action might be guaranteed further by a physical interaction between mtNOS-I and CcOX. This possibility has never been investigated. Here we demonstrate that mtNOS-I is associated with CcOX, as proven by electron microscopic immunolocalization and co-immunoprecipitation studies. By affinity chromatography, we found that association is due to physical interaction of mtNOS-I with the C-terminal peptide of the Va subunit of CcOX, which displays a consensus sequence for binding to the PDZ domain of mtNOS-I previously unreported for CcOX. The molecular details of the interaction have been analyzed by means of molecular modeling and molecular dynamics simulations. This is the first evidence of a protein-protein interaction mediated by PDZ domains involving CcOX.     

Interchangeability and tolerability of a virosomal and an aluminum-adsorbed hepatitis A vaccine

The interchangeability of virosomal (Epaxal) and aluminum-adsorbed (Havrix 1440) hepatitis A virus (HAV) vaccines was studied in 111 healthy adults who were vaccinated in a randomized, single-blind, crossover clinical trial. Anti-HAV antibody titers were measured at days 0 (first dose), 14, and 28, and months 3, 6, 12 (second dose), 13, 24, 36, 48, 60 and 72. Most subjects (>95%) had sero-converted 14 days after the first dose of either vaccine. The second dose with either vaccine induced a high antibody response in all vaccines, irrespective of the type of vaccine administered as the first dose. Although both vaccines were well tolerated, the incidence of local adverse events (in particular pain) was significantly lower in subjects receiving the virosomal vaccine. Six-year follow-up data did not reveal any significant differences between the vaccination groups.     

Cotinine and N-(2-hydroxyethyl)valine as markers of passive exposure to tobacco smoke in children

Large segments of populations, including children, are exposed to environmental tobacco smoke (ETS), a risk factor for lung cancer and heart, circulatory and respiratory diseases. Recently, ETS was classified as a class A carcinogen by USEPA, as carcinogenic to humans by IARC (group 1) and by the National Toxicology Program of the US National Institutes of Health. Cotinine, a product of the metabolism of nicotine, is measurable in urine and, correlates strictly and directly to ETS exposure, therefore representing a well-known internal dose marker. Another marker of active tobacco smoking is the N-(2-hydroxyethyl) valine (HOEtVal) which results from the reaction between ethylene oxide (EtO) and the N-terminal valine of hemoglobin. The aim of this study was the evaluations of ETS markers, namely urinary cotinine and HOEtVal measured in blood in 100 children with ages ranging between 3 and 13 years. Experimental findings show that cotinine, as a specific internal dose marker, and HOEtVal, as a nonspecific biological effective dose marker, both depend on the passive exposure to ETS as well as on the active habit of smoking.     

Green tea polyphenol extract attenuates ischemia/reperfusion injury of the gut

Various studies have clearly demonstrated that green tea catechins possess potent antioxidative properties, and the preventive effects against various oxidative diseases have been reported. The aim of this study was to investigate the effect of green tea extract on the tissue injury caused by ischemia/reperfusion (I/R) of the gut. I/R injury of the intestine was caused by clamping both the superior mesenteric artery and the celiac trunk for 45 min followed by release of the clamp allowing reperfusion for 1 h or 4 h. This procedure results in splanchnic artery occlusion (SAO) shock. Rats subjected to SAO developed a significant fall in mean arterial blood pressure, and only 10% of the animals survived for the entire 4-h reperfusion period. Surviving animals were sacrificed for histological examination and biochemical studies. Rats subjected to SAO displayed a significant increase in tissue myeloperoxidase (MPO) activity and malondialdehyde (MDA) levels, significant increases in plasma tumor necrosis factor (TNF)- levels and marked injury to the distal ileum. Increased immunoreactivity to nitrotyrosine was observed in the ileum of rats subjected to SAO. Staining of sections of the ileum obtained from SAO rats with anti-intercellular adhesion molecule (ICAM-1) antibody and with anti-P-selectin antibody resulted in diffuse staining. Administration of green tea extract (20 and 10 mg kg^–1 i.v.) 15 min prior to the onset of gut reperfusion significantly reduced in a dose-dependent manner the fall in mean arterial blood pressure, the mortality rate, infiltration of the reperfused intestine with polymorphonuclear neutrophils (MPO activity), lipid peroxidation (MDA levels), production of TNF-, and histological evidence of gut injury. Administration of green tea extract also markedly reduced nitrotyrosine formation and the up-regulation of ICAM-1 and P-selectin during reperfusion. In order to clarify that green tea extract might be useful in the therapy of I/R injury, we also investigated the effect of green tea extract (20 mg kg^–1 i.v.) when administered 5 min after the onset of gut reperfusion. Similar to the pretreatment approach, the post-treatment also significantly reduced the gut injury induced by I/R. These results demonstrate that green tea extract significantly reduces I/R injury of the intestine.