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Konrad A.,University Medical Center Erlangen
Methods in molecular biology (Clifton, N.J.) | Year: 2011

Several human pathogenic viruses encode large genomes with often more than 100 genes. Viral pathogenicity is determined by carefully orchestrated co-operative activities of several different viral genes which trigger the phenotypic functions of the infected cells. Systematic analyses of these complex interactions require high-throughput transfection technology. Here we have provided a laboratory manual for the reverse transfected cell microarray (RTCM; alternative name: cell chip) as a high-throughput transfection procedure, which has been successfully applied for the systematic analyses of single and combination effects of genes encoded by the human herpesvirus-8 on the NF-kappaB signal transduction pathway. In order to quantitatively determine the effects of viral genes in transfected cells, protocols for the use of GFP as an indicator gene and for indirect immunofluorescence staining of cellular target proteins have been included. RTCM provides a useful methodological approach to investigate systematically combination effects of viral genes on cellular functions.

Kallmunzer B.,Friedrich - Alexander - University, Erlangen - Nuremberg | Breuer L.,Friedrich - Alexander - University, Erlangen - Nuremberg | Kahl N.,Friedrich - Alexander - University, Erlangen - Nuremberg | Bobinger T.,Friedrich - Alexander - University, Erlangen - Nuremberg | And 4 more authors.
Stroke | Year: 2012

Background and Purpose-Patients with acute cerebrovascular events are susceptible to serious cardiac arrhythmias, but data on the time course and the determinants of their onset are scarce. Methods-The prospective Stroke-Arrhythmia-Monitoring-Database (SAMBA) assessed cardiac arrhythmias with need for urgent evaluation and treatment in 501 acute neurovascular patients during the first 72 hours after admission to a monitored stroke unit. Arrhythmias were systematically detected by structured processing of telemetric data. Time of arrhythmia onset and predisposing factors were investigated. Results-Significant cardiac arrhythmias occurred in 25.1% of all patients. Incidence was highest during the first 24 hours after admission. Serious arrhythmic tachycardia (ventricular or supraventricular >130 beats/min) was more frequent than bradycardic arrhythmia (sinus-node dysfunction, bradyarrhythmia, or atrioventricular block °II and °III). Arrhythmias were independently associated with higher age and severer neurological deficits as measured by the National Institutes of Health Stroke Scale on admission. Conclusions-The risk for significant cardiac arrhythmia after an acute cerebrovascular event is highest during the first 24 hours of care and declines with time during the first 3 days. Along with established vascular risk factors, the National Institutes of Health Stroke Scale may be considered for a stratified allocation of monitoring capabilities. Clinical Trial Registration-URL: www.clinicaltrials.gov. Unique identifier: NCT01177748. © 2012 American Heart Association, Inc.

Naschberger E.,University Medical Center Erlangen
Methods in molecular biology (Clifton, N.J.) | Year: 2011

Antiangiogenic drugs have been used successfully for the treatment of colorectal cancer (CRC) and several other tumor types. Until recently, viable tumor endothelial cells (TEC) and normal endothelial cells of uninvolved colon tissue (NEC) from the same patient have not been available to optimize treatment strategies in vitro. Here, we describe a protocol for the isolation of TEC and NEC. These cells were isolated at a very high purity via magnetic cell sorting of tissue samples obtained from surgical specimens of patients suffering from CRC. Isolated TEC and NEC expressed CD31, CD105, VE-cadherin, VCAM-1, ICAM-1, and E-selectin, formed capillaries in basal membrane extract, and were able to take up acetylated LDL. They were negative for podoplanin, CD45, CD68, and CK-20, indicating blood vessel endothelial lineage. Expression of vWF was more pronounced in NEC cultures, whereas vWF was absent or only slightly expressed in all TEC cultures in vitro. Lower intracellular concentrations of vWF were also detected in TEC as compared to NEC at the tissue level. The latter finding demonstrated that differential features of TEC and NEC in vivo are stably perpetuated in culture. The isolated endothelial cell cultures may provide a useful in vitro model system to elucidate epigenetic effects on angiogenesis in cancer and to optimize antiangiogenic therapy.

Kovacevic M.,HNO Praxis am Hanse Viertel | Wurm J.,University Medical Center Erlangen
Facial Plastic Surgery Clinics of North America | Year: 2015

Nasal hump excision is common during septorhinoplasty. Without appropriate restoration of the middle nasal vault, cosmetic and functional problems may ensue. Recently, spreader flaps have become an established alternative to traditional spreader grafts. Typical indications include primary rhinoplasty patients with hump noses, hump/tension noses, and moderately hooked or crooked noses. When suitable patients are selected, spreader flaps and their modifications represent a reliable alternative to the standard spreader graft, and when all of the necessary prerequisites are met, this technique obviates the need for additional cartilage grafting in most cases. © 2015 Elsevier Inc.

Naschberger E.,University Medical Center Erlangen | Wenzel J.,University of Bonn | Kretz C.C.,German Cancer Research Center | Herrmann M.,University Medical Center Erlangen | And 3 more authors.
Experimental Dermatology | Year: 2011

The large GTPase human guanylate binding protein-1 (GBP-1) is a key mediator of angiostatic effects of inflammation and is induced by interferon (IFN)-α and IFN-γ in endothelial cells (ECs). The aim of this study was to investigate whether GBP-1 is a marker of skin lesions in patients with cutaneous lupus erythematosus (CLE). Western blotting revealed that GBP-1 was in vitro induced by IFN-α and -γ in primary keratinocytes obtained from healthy controls. Moreover, we found that this protein was expressed by keratinocytes and ECs in primary and ultraviolet (UV)-induced skin lesions from patients with various subtypes of CLE, when compared to non-lesional skin. No GBP-1 expression was noted in skin biopsy specimens 24 or 72h after UV irradiation prior to lesion formation in patients with CLE or in healthy control specimens with or without UV irradiation. Initial findings suggest that GBP-1 is not expressed in other skin diseases with different inflammatory aetiology, such as atopic dermatitis. We conclude that GBP-1 expression is closely associated with skin lesions in patients with CLE, suggesting a contribution of GBP-1 in the pathogenesis of this disease. © 2011 John Wiley & Sons A/S.

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