Glycinergic neurons expressing enhanced green fluorescent protein in bacterial artificial chromosome transgenic mice

Although glycine is a major inhibitory transmitter in the mammalian CNS, the role of glycinergic neurons in defined neuronal circuits remains ill defined. This is due in part to difficulties in identifying these cells in living slice preparations for electrophysiological recordings and visualizing their axonal projections. To facilitate the morphological and functional analysis of glycinergic neurons, we generated bacterial artificial chromosome (BAC) transgenic mice, which specifically express enhanced green fluorescent protein (EGFP) under the control of the promotor of the glycine transporter (GlyT) 2 gene, which is a reliable marker for glycinergic neurons. Neurons expressing GlyT2-EGFP were intensely fluorescent, and their dendrites and axons could be visualized in great detail. Numerous positive neurons were detected in the spinal cord, brainstem, and cerebellum. The hypothalamus, intralaminar nuclei of the thalamus, and basal forebrain also received a dense GlyT2-EGFP innervation, whereas in the olfactory bulb, striatum, neocortex, hippocampus, and amygdala positive fibers were much less abundant. No GlyT2-EGFP-positive cell bodies were seen in the forebrain. On the subcellular level, GlyT2-EGFP fluorescence was colocalized extensively with glycine immunoreactivity in somata and dendrites and with both glycine and GlyT2 immunoreactivity in axon terminals, as shown by triple staining at all levels of the neuraxis, confirming the selective expression of the transgene in glycinergic neurons. In slice preparations of the spinal cord, no difference between the functional properties of EGFP-positive and negative neurons could be detected, confirming the utility of visually identifying glycinergic neurons to investigate their functional role in electrophysiological studies.     

Serum antibodies against the hepatitis C virus E2 protein mediate antibody-dependent cellular cytotoxicity (ADCC)

BACKGROUND/AIMS: The role of antibody dependent cellular cytotoxicity (ADCC) in HCV infection is unclear at present. Antibodies mediating ADCC are usually directed against viral envelope proteins. As cell surface expression of the HCV envelope E2 protein has been shown, the HCV E2 protein is an especially promising candidate target for ADCC. METHODS: Sera from patients with acute (n=6), self-limited (n=11) and chronic (n=19) HCV infection were analyzed in this study. Sera reacting with cell-bound HCV antigens were examined in a flowcytometric cytotoxicity assay using antigen-coated JOK-1 cells as targets. RESULTS: We found that sera from all stages of HCV infection reacted with cells loaded with HCV E2. E2-specific ADCC was observed in patients with acute (n=3/6), self-limited (n=5/11) and chronic (n=13/19) hepatitis C and was closely related to fluorescence intensity in the E2-binding assay (r=0.67, P<0.001). CONCLUSIONS: We conclude that E2-antibodies from all stages of HCV infection can mediate ADCC. Thus, the role of this process in the pathogenesis of chronic hepatitis C should be further elucidated.     

Sustained, spontaneous disappearance of serum HCV-RNA under immunosuppression after liver transplantation for HCV cirrhosis

Immunosuppression is a main determinant for the increased Hepatitis C Virus (HCV) replication after liver transplantation and the accelerated course of recurrent HCV liver disease. We present two patients both with diabetes, renal dysfunction with proteinuria converted to sirolimus therapy, who cleared serum HCV RNA without antiviral treatment. This is a potentially important observation that should stimulate study into factors that may help viral clearance from blood.     

Epidemiology, risk factors, and genetics of high-altitude-related pulmonary disease

High-altitude-related pulmonary disease is a spectrum of acute and chronic illnesses with a well-described epidemiology. The risk for these illnesses is related to well-known environmental risk factors and lesser-known but important genetic factors. Prevention of acute high-altitude illness is possible in most visitors from lower elevations. Chronic high-altitude illnesses have an important worldwide impact.     

Magnetic resonance imaging gastrography: evaluation of the dark lumen technique compared with conventional gastroscopy in patients with malignant gastric disease

OBJECTIVES: We sought to demonstrate the feasibility of depicting gastric tumors using magnetic resonance imaging (MRI) while applying the dark lumen technique. The findings were correlated with conventional gastroscopy. In addition, we evaluated the screening for lymph nodes and liver metastases during the same session to identify potential tumor spread. MATERIALS AND METHODS: The study included 15 patients with known malignant gastric disease. Conventional gastrointestinal endoscopy was performed in all patients as gold standard. All patients were examined with computed tomography for tumor staging. MRI was performed using 1 L of tap water as oral contrast agent for all protocols. The MRI program included an axial T1-weighted (T1w; 2D-FLASH) sequence, an axial STIR and T2w (TSE) sequence, and 2 postcontrast T1w (3D coronal /2D axial FLASH) sequences using 0.2 mmol/kg gadolinium diethylenetriaminepenta-acetic acid as intravenously injected contrast agent. Qualitative analysis and comparison with conventional gastroscopy were performed. RESULTS: The images obtained with the postcontrast 3D coronal T1w-FLASH sequence were the most suitable in identifying gastric tumors. Complete correlation with conventional gastroscopy was achieved in 80% of the cases and partial correlation in 13% of the cases. The same imaging sequence was also appropriate for the evaluation of lymph nodes. For the identification of liver metastases, the images obtained with the axial postcontrast T1w 2D-FLASH sequence provided a higher diagnostic confidence as compared with other imaging protocols. CONCLUSIONS: Applying the dark lumen technique through MRI is suitable for imaging gastric tumors and has the potential to become a "one-stop shopping" method because of the possibility for lymph node evaluation and screening for metastases during the same session based on the same images.     

Qualität digitaler Mammogramme

Digital imaging in mammography is becoming more and more accepted using both computed (CR) and direct radiography (DR). These techniques will soon be used in screening programs. Therefore, quality assurance for this technique is indispensable. The relevance of the current regulations, such as EPOC and the German QS-RL was investigated. For the investigation, a breast phantom and appropriate software were developed. Both were tested using digital mammography systems from six manufactures. Quality assurance parameters (such as contrast to noise ratio and contrast resolution) were calculated from these data sets. The results should be considered in future standards for mammography (IEC respectively DIN). In addition, this type of test procedure is time saving and enables a reduction in test devices, i. e. in costs.