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Conventional panoramic radiography (cPR) underlines procedure-related limitations in the display of objects. CBCT is presumed to overcome these constraints. To virtualize a cPR view, reformatted panoramic images (rPIs) can be generated. This study evaluated the rPI with regard to its susceptibility to sterical object deposition in comparison with cPR.A specially developed implant model with dental implants each of 4.0-mm diameter and 11.0-mm length was depositioned by shift, rotation and tilt of 5.00 mm (±0.01 mm) of horizontal shift and 5.0° (±0.167°), respectively, on a highly precise goniometer rotation table, and cPRs and rPIs were generated. Automated evaluation of the cPRs was carried out using a specially developed software. rPIs were processed and analyzed by a semi-automated image analysis.Object deposition lead to distortive effects in the rPI analogue to cPR, but they appear in display only. Objects illustrated in the rPI were dimensionally correct, but sterical relations are elusive. Results are obtained for the horizontal shift, declination and reclination, lateral tilt and rotation.Distortions within the rPI represent the illustration of the hyperbolic-shaped layer out of the three-dimensional data set. With this study, we demonstrated these procedure-related inherent but practically underestimated consequences. Effects of sterical object malpositioning must be compensated by the observer by adequate virtual adjustment of the processed layer. Accurate virtual adjustment leads to vertical dimensions. Sterical relations, e.g. angulation of two objects, are irretraceable unless precisely referenced.

Throughout the last years, the contribution of alveolar type II epithelial cells (AECII) to various aspects of immune regulation in the lung has been increasingly recognized. AECII have been shown to participate in cytokine production in inflamed airways and to even act as antigen-presenting cells in both infection and T-cell mediated autoimmunity (1-8). Therefore, they are especially interesting also in clinical contexts such as airway hyper-reactivity to foreign and self-antigens as well as infections that directly or indirectly target AECII. However, our understanding of the detailed immunologic functions served by alveolar type II epithelial cells in the healthy lung as well as in inflammation remains fragmentary. Many studies regarding AECII function are performed using mouse or human alveolar epithelial cell lines (9-12). Working with cell lines certainly offers a range of benefits, such as the availability of large numbers of cells for extensive analyses. However, we believe the use of primary murine AECII allows a better understanding of the role of this cell type in complex processes like infection or autoimmune inflammation. Primary murine AECII can be isolated directly from animals suffering from such respiratory conditions, meaning they have been subject to all additional extrinsic factors playing a role in the analyzed setting. As an example, viable AECII can be isolated from mice intranasally infected with influenza A virus, which primarily targets these cells for replication (13). Importantly, through ex vivo infection of AECII isolated from healthy mice, studies of the cellular responses mounted upon infection can be further extended. Our protocol for the isolation of primary murine AECII is based on enzymatic digestion of the mouse lung followed by labeling of the resulting cell suspension with antibodies specific for CD11c, CD11b, F4/80, CD19, CD45 and CD16/CD32. Granular AECII are then identified as the unlabeled and sideward scatter high (SSC(high)) cell population and are separated by fluorescence activated cell sorting (3). In comparison to alternative methods of isolating primary epithelial cells from mouse lungs, our protocol for flow cytometric isolation of AECII by negative selection yields untouched, highly viable and pure AECII in relatively short time. Additionally, and in contrast to conventional methods of isolation by panning and depletion of lymphocytes via binding of antibody-coupled magnetic beads (14, 15), flow cytometric cell-sorting allows discrimination by means of cell size and granularity. Given that instrumentation for flow cytometric cell sorting is available, the described procedure can be applied at relatively low costs. Next to standard antibodies and enzymes for lung disintegration, no additional reagents such as magnetic beads are required. The isolated cells are suitable for a wide range of functional and molecular studies, which include in vitro culture and T-cell stimulation assays as well as transcriptome, proteome or secretome analyses (3, 4).

Die plastisch-rekonstruktive Chirurgie im Bereich des Gesichtes hatte schon immer den Anforderungen der Wiederherstellung von Aussehen und Funktion zu genügen, damit Patienten mit tumor- oder traumabedingten Defekten oder angeborenen Fehlbildungen posttherapeutisch erfolgreich rehabilitiert werden können. Um dieses Ziel zu erreichen, kommen derzeit drei wesentliche Techniken zum Einsatz: lokale Nahlappenplastiken, gestielte regionale Lappen und mikrochirurgische Transplantate. Die vorliegende Übersichtsarbeit beschreibt diese drei Haupttechniken der plastisch-rekonstruktiven Chirurgie des Gesichtes und gibt einen Ausblick auf neuartige Technologien wie der virtuellen Planung von rekonstruktiven Eingriffen., GMS Interdisciplinary Plastic and Reconstructive Surgery DGPW; 1:Doc12; ISSN 2193-8091

This report describes physical and imaging findings in a case of anterior lingual mandibular bone depression in a child. This entity is very rarely diagnosed and even more extremely rarely seen in children. We present some characteristic findings depicted on images provided by different sources and briefly address current hypotheses on its pathogenesis.

The differential diagnosis of osseous jaw lesions includes numerous entities. A specific mandibular bone defect known as Stafne's cavity or cyst presents wirh a characteristic cyst-like lesion on radiographs of the jaw. Although the differential diagnosis from other jaw lesions is mandatory, this lesion does not usually require surgical therapy. Current theories concerning the pathogenesis of Stafne's cavity prefer bone depression as a consequence of constant pressure arising from an adjacent salivary gland. We present a case of a large Stafne's cavity eroding the basal mandibular cortex that was found accidentally on an orthopantogram taken for diagnosis of dental diseases. The patient noted a slight pain on pressure during physical investigation. The lesion was completely occupied by fatty tissue, as shown during surgical exploration and as revealed by histological investigation of the specimen. The theory of parenchymal pressure as the cause of mandibular bone depression appears to be inapplicable in the present case. The pathogenesis of Stafne's cavity is still obscure. Differential diagnosis of mandibular lesions is essential in adequate treatment planning.

Analyzing chromosomal amplifications delivers valuable information for identification of oncogenes. For carcinomas of the oral cavity only few genes have been identified in amplified regions. The aim of this study was to search genes in amplified regions as possible biomarkers and targets for novel therapies.DNA from 10 carcinomas of the floor of the oral cavity was examined using a 500K Array GeneChip (Affymetrix 6.0) to detect chromosomal losses, gains or amplifications. Suspicious alterations were validated on tissue microarrays using fluorescence in situ hybridization (FISH) with respective probes.FISH-validation on tissue arrays confirmed PPFIA1 amplifications as one of the most frequent events (32.6%). High (10-20 signals) and low (10 signals) amplification of PPFIA1 was found in 10.9% (5/46) and 21.7% (10/46) tumours, respectively. Fine mapping with overlapping FISH probes showed co-amplification of PPFIA1 and the Cyclin D1 gene which are approximately 600 kb apart from each other, likely in the same amplicon.PPFIA1 was frequently co-amplified with the Cyclin D1 gene in oral carcinomas and could present a biomarker as well as a novel target for specific gene therapy. Further studies are necessary to investigate the role of PPFIA1 in development and pathogenesis of oral carcinomas.

Keratocystic odontogenic tumors (KCOTs) arise from remnants of epithelial structures associated with the development of teeth and occur predominantly intraosseously. In rare occasions they can appear extraosseously in the gingiva as peripheral counterparts; only 15 cases have been reported to date. Evenmore rare are cases of KCOTs of the buccal soft tissues. The aim of this report is to present a rare case of KCOT affecting the buccal soft tissue as an original site and a review of the literature regarding diagnostic and therapeutic options.

The present study aims to investigate the relevance of immunohistochemical p53 expression in carcinomas of the oral cavity and of the head and neck region. Long-term clinical and histopathological follow-up findings as well as HPV status are correlated with the results of this examination.Sections made from two tissue arrays composed of 222 oral squamous cell carcinomas and 427 squamous cell carcinomas of the head and neck region, respectively, were examined for p53 expression and Ki-67 index by means of immunohistochemistry. Correlation of long-term clinical findings of the patients and pathological features of tumours with laboratory results were examined statistically.No significant correlation was found between the p53 immunohistochemical expression in the 207 oral carcinomas and features of the tumours and patient outcomes. There was no significant association between the Ki-67 labelling index and the p53 expression.Our failure in detecting any association of the p53 immunohistochemical expression regardless of HPV status with clinical features of these tumours suggests it lacks a prognostic value for squamous cell carcinomas of the oral cavity.The prognostic value of p53 immunostaining in oral squamous carcinoma is not clarified yet. In the present study, there is no impact on any prognostical item nor even a correlation with cell proliferation (Ki-67) regardless of HPV status.

The question which bacterial species are present in water and if they are viable is essential for drinking water safety but also of general relevance in aquatic ecology. To approach this question we combined propidium iodide/SYTO9 staining ("live/dead staining" indicating membrane integrity), fluorescence-activated cell sorting (FACS) and community fingerprinting for the analysis of a set of tap water samples. Live/dead staining revealed that about half of the bacteria in the tap water had intact membranes. Molecular analysis using 16S rRNA and 16S rRNA gene-based single-strand conformation polymorphism (SSCP) fingerprints and sequencing of drinking water bacteria before and after FACS sorting revealed: (1) the DNA- and RNA-based overall community structure differed substantially, (2) the community retrieved from RNA and DNA reflected different bacterial species, classified as 53 phylotypes (with only two common phylotypes), (3) the percentage of phylotypes with intact membranes or damaged cells were comparable for RNA- and DNA-based analyses, and (4) the retrieved species were primarily of aquatic origin. The pronounced difference between phylotypes obtained from DNA extracts (dominated by Betaproteobacteria, Bacteroidetes, and Actinobacteria) and from RNA extracts (dominated by Alpha-, Beta-, Gammaproteobacteria, Bacteroidetes, and Cyanobacteria) demonstrate the relevance of concomitant RNA and DNA analyses for drinking water studies. Unexpected was that a comparable fraction (about 21%) of phylotypes with membrane-injured cells was observed for DNA- and RNA-based analyses, contradicting the current understanding that RNA-based analyses represent the actively growing fraction of the bacterial community. Overall, we think that this combined approach provides an interesting tool for a concomitant phylogenetic and viability analysis of bacterial species of drinking water.

The aromatic peroxygenase (APO; EC 1.11.2.1) from the agraric basidomycete Marasmius rotula (MroAPO) immobilized at the chitosan-capped gold-nanoparticle-modified glassy carbon electrode displayed a pair of redox peaks with a midpoint potential of -278.5 mV vs. AgCl/AgCl (1 M KCl) for the Fe(2+)/Fe(3+) redox couple of the heme-thiolate-containing protein. MroAPO oxidizes aromatic substrates such as aniline, p-aminophenol, hydroquinone, resorcinol, catechol, and paracetamol by means of hydrogen peroxide. The substrate spectrum overlaps with those of cytochrome P450s and plant peroxidases which are relevant in environmental analysis and drug monitoring. In M. rotula peroxygenase-based enzyme electrodes, the signal is generated by the reduction of electrode-active reaction products (e.g., p-benzoquinone and p-quinoneimine) with electro-enzymatic recycling of the analyte. In these enzyme electrodes, the signal reflects the conversion of all substrates thus representing an overall parameter in complex media. The performance of these sensors and their further development are discussed.

We used bacterial cellulose membranes as outer membranes for glucose sensors based upon amperometric detection of glucose oxidase reaction products. We found a long-term stability in 1:10 diluted blood of about 200 h. In comparison a Cuprophan® membrane was stable for only 30 h under the same conditions. Comparing the two membranes in undiluted human blood, a similar trend was found in the long-term stability: Cuprophan® was stable for only 3–4 h and the glucose sensor with the bacterial cellulose membrane was stable for more than 24 h. In addition, the measuring range of the glucose sensor could be extended up to 170 mM glucose by treating the bacterial cellulose membrane with a polyamide.

Einleitung: Die Regeneration von Knorpeldefekten stellt in der Klinik ein Problem dar. Sowohl Knorpeltransplantate als auch im Labor generierte Knorpelkonstrukte weisen eine starke Degeneration unter Belastung auf. Häufig mangelt es vor Ort an einer ausreichenden regenerativen Eigenkapazität[for full text, please go to the a.m. URL], 49. Jahrestagung der Deutschen Gesellschaft für Plastische und Wiederherstellungschirurgie (DGPW)

Type 1 neurofibromatosis (NF-1), also known as von Recklinghausen disease, is caused by a disorder of a single gene on chromosome 17 that usually restrains cell division. A sequence that is frequently associated with NF-1 is tumor progression from neurofibromas to malignant peripheral nerve sheath tumors (MPNSTs). The aim of this study was to determine the expression of the neural L1 cell adhesion molecule in dermal-diffuse neurofibromas, plexiform neurofibromas, and MPNSTs of NF-1. We retrospectively analyzed surgically resected primary tumors, including 20 dermal neurofibromas, 23 plexiform neurofibromas, and 17 MPNSTs, by immunohistochemistry in paraffin sections of NF-1 tumors with the use of the L1-specific monoclonal antibody UJ127, which does not cross-react with other members of the L1 family. Immunostainings for CD34 and S100 were included to distinguish and allocate L1-expressing Schwann cells and perineural (specialized) fibroblasts. Our data showed that L1 is highly expressed in all benign NF-1 tumors and in some but not all MPNSTs. Furthermore, we demonstrated a correlation between L1 expression and differentiation grade of MPNSTs. There was a significant trend toward lower or nondetectable expression in the poorly differentiated MPNSTs, in contrast to all other tumor entities so far investigated, in which L1 expression correlated positive with malignancy, except for juvenile but not adult-derived neuroblastomas. Future studies are warranted to elucidate the molecular basis of the varying effects of the degree of L1 expression, receptor, and signal transduction mechanisms in different tumors.

Einleitung: Die Rekonstruktion des Unterkiefers mit mikrovaskulären Fibulatransplantaten nach Resektion neoplastischer Raumforderungen erfordert einen umfangreichen Eingriff zur möglichst weitgehenden Wiederherstellung von Funktion und Ästhetik. Um diese hohen Anforderungen zu erfüllen[for full text, please go to the a.m. URL], 49. Jahrestagung der Deutschen Gesellschaft für Plastische und Wiederherstellungschirurgie (DGPW)

Background Surveys from the USA, Australia and Spain have shown significant inter-institutional variation in delivery room (DR) management of very low birth weight infants (VLBWI, < 1500 g) at birth, despite regularly updated international guidelines. Objective To investigate protocols for DR management of VLBWI in Germany, Austria and Switzerland and to compare these with the 2005 ILCOR guidelines. Methods DR management protocols were surveyed in a prospective, questionnaire-based survey in 2008. Results were compared between countries and between academic and non-academic units. Protocols were compared to the 2005 ILCOR guidelines. Results In total, 190/249 units (76%) replied. Protocols for DR management existed in 94% of units. Statistically significant differences between countries were found regarding provision of 24 hr in house neonatal service; presence of a designated resuscitation area; devices for respiratory support; use of pressure-controlled manual ventilation devices; volume control by respirator; and dosage of Surfactant. There were no statistically significant differences regarding application and monitoring of supplementary oxygen, or targeted saturation levels, or for the use of sustained inflations. Comparison of academic and non-academic hospitals showed no significant differences, apart from the targeted saturation levels (SpO2) at 10 min. of life. Comparison with ILCOR guidelines showed good adherence to the 2005 recommendations. Summary Delivery room management in German, Austrian and Swiss neonatal units was commonly based on written protocols. Only minor differences were found regarding the DR setup, devices used and the targeted ranges for SpO2 and FiO2. DR management was in good accordance with 2005 ILCOR guidelines, some units already incorporated evidence beyond the ILCOR statement into their routine practice.

Purpose Corrective plastic surgery is indicated after accidents, burns, cancer surgery, or postoperative wound healing disorders with large tissue defects. The range of reconstructive techniques includes local skin flaps, pedicle grafts, and microvascular anastomosed flaps in the case of large defects. Main complications such as arterial and venous insufficiency caused by a vessel collapse or a vascular spasm are reported regularly in the area of anastomosed vessels and are the concern of any surgeon. Today, leeches are used if wound healing is at risk because of hemodynamic imbalance or a venous insufficiency. Patients and Methods A retrospective evaluation of 148 patients who underwent medical leech therapy in the case of local or pedicaled flaps and some patients who had undergone reconstruction with microvascular flaps from 2005 and 2010 was conducted. Our sample had the typical symptoms of venous congestions of their flaps, despite suture removal, relief of pressure on the flap, and the elimination of a hematoma beyond the flap after surgery. Medical leech therapy was used in these cases. Results Our series has confirmed the excellent and predictable healing after medical leech therapy for local and microsurgical anastomosed flaps in the case of venous congestion. Conclusion Leech therapy should be considered as a reliable additional procedure and an advantage in maxillofacial and plastic reconstructive surgery to remedy complications resulting from a hemodynamic imbalance or venous insufficiency in the immediate postoperative period.

Complications can result from the surgical removal of third molars, and the complication rate is expected to be higher in immunosuppressed patients. However, such cases are rare. Leflunomide is a slow-acting antirheumatic agent used in the treatment of rheumatoid arthritis and psoriatic arthritis. It inhibits dihydroorotate dehydrogenase, a key enzyme in pyrimidine metabolism that is particularly significant in the cell division of T lymphocytes, and suppresses leukocyte migration. In the intestinal wall and the liver, leflunomide is converted into its active metabolite, A771726, which is further metabolized into additional active substances. The half-life of these active leflunomide metabolites is between 1 and 4 weeks. Known side effects of leflunomide are itching, mucous membrane lesions, hair loss (alopecia), diarrhea, and very rarely, agranulocytosis and fatal liver necrosis. Combination therapies with leflunomide and other basic antirheumatic drugs such as methotrexate and chloroquine are more effective than monotherapy and are widely used in nonsurgical rheumatology. However, the combination of leflunomide and methotrexate is associated with the risk of severe liver damage (eg, granulomatous hepatitis) and/or lifethreatening infections. Furthermore, the risks of longterm treatment with such a combination therapy are not yet well-known. We report for the first time a case of life-threatening complications after surgical removal of the infected mandibular third molar in a patient with leflunomideinduced pancytopenia.

Bei der Koagulation von phasenbildenden, unvertraglichen Polymermischungslosungen entstehen durch das Zusatzpolymer bedingte, bemerkenswert hochporose Fallungsstrukturen. Die dargestellten Untersuchungen haben das Ziel, einen moglichen Einflus der Zusatzpolymere auf die Porositat der Fallungsprimarschicht zu erfassen. Anhand von Permeationsuntersuchungen konnte an den auf fester Unterlage hergestellten Flachmembranen im Gegensatz zur Hohlmembran eine erhohte Porositat der Fallungsprimarschicht nachgewiesen werden. Hierfur wird ein Porenbildungsmechanismus vorgeschlagen. Remarkable highly porous structures are formed by the coagulation of incompatible phase-forming polymer blend solutions. The effect of an added polymer on the porosity of the primary layer is studied. Investigations of the permeability of membranes prepared by different procedures revealed that only in flat membranes prepared on a solid support an elevated porosity of the primary coagulated layer could be observed. A mechanism of pore formation in this special case is suggested.

Bei der Koagulation von phasenbildenden, unvertraglichen Polymermischungslosungssystemen entstehen hochporose Fallungsstrukturen. Das maximal realisierbare Porenvolumen derart hergestellter Formkorper wird im wesentlichen durch das Volumen der dispergierten Zusatzpolymerlosungsphase in der Polymermischungslosung bestimmt. Wie am Beispiel des Polymermischungslosungssystems PAN/SAN/DMF gezeigt wird, gilt dieses Strukturbildungsprinzip auch im allgemeinen Mischungssystem Polymer(A)/Copolymer(AB)/Losungsmittel, wenn sowohl die Solvatationsverhaltnisse in der Mischungslosung als auch Veranderungen im Vertraglichkeitsverhalten berucksichtigt werden. Highly porous coagulated structures are formed by coagulating incompatible phase-forming polymer blend solutions. The maximum of the pore volume in the so formed shaped bodies is essentially predetermined by the volume of the dispersed phase owned by the solution of the added polymer in the polymer blend solution. Taken as an example, the blend system PAN/SAN/DMF revealed this principle of structure formation also to be valid for the general system polymer(A)/copolymer(AB)/solvent by taking into consideration the relations of solvatation in the mixed solutions and variations of the compatibility relations, too.

IL-10 plays a central role in restraining the vigor of inflammatory responses, but the critical cellular sources of this counter-regulatory cytokine remain speculative in many disease models. Using a novel IL-10 transcriptional reporter mouse, we found an unexpected predominance of B cells (including plasma cells) among IL-10 expressing cells in peripheral lymphoid tissues at baseline and during diverse models of in vivo immunological challenge. Use of a novel B cell-specific IL-10 knockout mouse revealed that B cell-derived IL-10 non-redundantly decreases virus-specific CD8+ T cell responses and plasma cell expansion during murine cytomegalovirus (MCMV4) infection and modestly restrains immune activation after challenge with foreign antibodies to IgD. In contrast, no role for B cell-derived IL-10 was evident during endotoxemia; however, while B cells dominated lymphoid tissue IL-10 production in this model, myeloid cells were dominant in blood and liver. These data suggest that B cells are an under-appreciated source of counter-regulatory IL-10 production in lymphoid tissues, provide a clear rationale for testing the biological role of B cell-derived IL-10 in infectious and inflammatory disease, and underscore the utility of cell type-specific knockouts for mechanistic limning of immune counter-regulation.

Polyacrylonitrile membranes are prepared using three different membrane preparation principles. Characterization of the structure of the skin layer is carried out in more detail by parameters of separation curves and their pore size distribution function, the former being based on a logarithmic normal distribution. Under constant conditions of measurement, comparing considerations are reflected about the structure of the separating active layer of PAN bicomponent membranes and two differently prepared PAN monocomponent membranes. The parameters of the separation curves and pore size distributions are discussed. Polyacrylnitril-Membranen werden nach drei unterschiedlichen Membranbildungsprinzipien hergestellt. Die nahere Charakterisierung der Struktur der trennaktiven Schichten erfolgt anhand von Parametern der Trennkurven und der daraus berechneten Porengrosenverteilungen, wobei eine logarithmische Normalverteilung zugrunde gelegt wird. Bei konstanten Mesbedingungen werden vergleichende Betrachtungen der strukturellen Besonderheiten der trennaktiven Begrenzungsschicht von PAN-Bikomponentenmembranen und zwei unterschiedlich hergestellten PAN-Monokomponentenmembranen angestellt und anhand von Parametern der Trennkurven oder der Porengrosenverteilung diskutiert.

Die Sialylierung von Glykokonjugaten ist essentiell für eine Vielzahl von Prozessen in und zwischen Zellen. Neben Zelldifferenzierungen, Entzündungsreaktionen, Zell-Adhäsionen und Pathogenesen ist die Beeinflussung der Stabilität von Glykoproteinen im Serum von Bedeutung. Die Abspaltung der terminalen Sialinsäuren durch Sialidasen ermöglicht die Bindung an den Asialoglykoprotein-Rezeptor und der Abbau des Glykoproteins wird eingeleitet. Durch die gezielte Modifikation einzelner Monosaccharide sollte die Aktivität der Sialidasen beeinflusst und im Vergleich zu nativen Oligosacchariden bestimmt werden. Nach der Supplementation der Zelllinien CHO (murin), sowie HEK293 und K-562 (human) mit 2-Desoxy-Galactose konnte ein konzentrationsabhängiger Austausch gegen die subterminale Galactose durch HPLC- und MS-Methoden nachgewiesen werden. Der Anteil von 62 % 2-Desoxy-D-Galactose in CHO-Zellen führte zu einer fast vollständigen Resistenz gegenüber der unspezifischen Sialidase A. In den humanen Zelllinien wurden Einbauraten zwischen 23 und 25% festgestellt. Während die K-562-Zellen im Vergleich zu nicht-modifizierten Glykanen eine 1,6fach erhöhte Sialidase- Resistenz aufwiesen, war diese in den HEK293-Zellen erniedrigt. Generell erfolgte in Anwesenheit der 2-Desoxy-D-Galactose eine reduzierte Glykosylierung. Die terminale Präsenz nicht-natürlicher Sialinsäuren in Glykanen hatte keine Änderung des Glykan-Profils zur Folge. Die an der Acetyl- Gruppe modifizierten N-Acetylmannosamine wurden entsprechend dem nativen Vorläufer zu neuen Neuraminsäuren umgesetzt und erreichten in den Glykoproteinen der Zellen einen Anteil von 45-92%. Die höchsten Sialidase C-Resistenzen in der murinen (CHO) und humanen Zelllinie (HEK293) konnten für die Analoga N-Butanoyl- und N-Pentanoylmannosamin festgestellt werden, die gleichzeitig die geringsten Einbauraten vorwiesen. Die Menge an messbaren sialylierten Strukturen nach der Enzymbehandlung erhöhte sich um das maximal 2,5fache (CHO) bzw. 1,6fache (HEK293) in Anwesenheit artifizieller Neuraminsäuren. Somit stellen beide Modifikationen, 2dGal und N-Acylmannosamine, eine Möglichkeit zur Verlängerung der Halblebenszeit von Glykoproteinen dar. Die Optimierung der Massenspektrometrie-Methodik und die Etablierung der Fragmentierung mittels Ionenfallen-Technik ermöglichte die Glykan-Charakterisierung ohne Fluoreszenz-Markierung mit HPLC-Auftrennungen. Die Auswertung der erhaltenen MS-Daten wurde mit Hilfe von online verfügbaren Programmen standardisiert. Für die Kalkulation der molekularen Massen von Glykanen, Peptiden und Glykoproteinen war der Aufbau einer Datenbank- basierenden Anwendung erforderlich. Das resultierende GlycoProtMass ermöglichte sowohl die Berechnungen von Molekülmassen, als auch deren Archivierung mit zusätzlichen Informationen., The sialylation of glycoconjugates is important for many cellular processes in cells and cell-cell contacts. Besides cell differentiation, inflammation, cell adhesion and pathogenesis, sialic acids influence the stability of glycoproteins. If these terminal monosaccharides are cleaved by sialidases glycoproteins bind to the asialoglycoprotein receptor and are degradated. The selective modification of monosaccharides should be tested for their influences on sialidase activities in comparison to native oligosaccharides. The supplementation of CHO cells (murine) and HEK293, K-562 (both human) with 2-deoxy-d-galactose (2dGal) led to an exchange against subterminal galactose. In CHO cells the glycans of membrane proteins had 62% 2-deoxy-galactoses incorporated in a nearly complete sialidase A resistance. The human cell lines showed incorporation rates of 23-25% for 2dGal. In comparison to unmodified oligosaccharides a 1,6-fold increased resistance was determined in K-562. In the case of HEK293 glycans were more sensitive against sialidase A treatment. Generally 2dGal induced a reduction in glycosylation. The also generated non- native sialic acids did not change the glycan profiles but the behaviour of these terminal monosaccharides. N-Acetylmannoseamines with modifications in the acetyl-group were used like native precursors of sialic acids and the resulting new neuraminic acids reached incorporation rates of 45-92%. The highest sialidase C resistance was detected for N-butanoyl- and N-pentanoylmannoseamine which also have shown the lowest incorporation rates. The new artificial monosaccharides led to an 2,5-fold higher sialidase resistance for CHO and 1,6-fold higher for HEK293 respectively. Both modifications, 2dGal and N-Acylmannoseamines, are good possibilities to modify the lifetime of glycoproteins. Optimized masspectrometric methods and established fragmentation with iontrap techniques were used to characterize glycans without fluorescence labelling with HPLC separations. The Analysis of resulting data was standardised with programs which are freely available in the internet. For the calculation of molecular masses of glycans, peptides and glycoproteins a new tool GlycoProtMass was developed. This program can be used simultaneously to calculate masses of molecules and to save results with additional information.

Der Einflus der Polymerlosungsschichtedickenverhaltnisse auf die Strukturbildung an der Innenschicht von Bikomponentenmembranen, die mit Hilfe einer Bikomponentenbreitschlitzduse hergestelt werden, wird untersucht. Anhand der Stofftransportdaten durch die Membran und durch rasterelektronenmikroskopische Untersuchungen werden mechanistische Vorstellungen zum Ablauf des Koagulationsgeschchens entwickelt. Die quantitative Beschreibung dieser Vorgange erfolgt mit Hilfe einer Losung des 2. Fickschen Gesetzes. Es werden scheinbare Diffusionkoeffizienten berechnet. Ein daraus ableitbarer Strukturubergangspunkt fur das System CA/DMF-PUR/DMF konnte experimentell bestatigt werden. The effect of the ratio of the thickness of the polymer solution layers on the structure formation at the inner layer of bicomponent membranes prepared by means of a bicomponent slit die is studied. Basing on the data of mass transfer through the membrane and REM investigations a conception of the coagulation process is suggested. The process is quantitatively described by the second Fick's law. Apparent diffusion coefficients are calculated. The structure transition point of the system CA/DMF-PUR/DMF has been derived from the results and was experimentally confirmed.

Serum response factor (SRF), is a crucial transcription factor for murine embryonic development and for the function of muscle cells and neurons. Gene expression data show that SRF and its transcriptional cofactors are also expressed in lymphocyte precursors and mature lymphocytes. However, the role of SRF in lymphocyte development has not been addressed in vivo so far, attributed in part to early embryonic lethality of conventional Srf-null mice. To determine the in vivo role of SRF in developing lymphocytes, we specifically inactivated the murine Srf gene during T or B cell development using lymphocyte-specific Cre transgenic mouse lines. T cell-specific Srf deletion led to a severe block in thymocyte development at the transition from CD4/CD8 double to single positive stage. The few residual T cells detectable in the periphery retained at least one functional Srf allele, thereby demonstrating the importance of SRF in T cell development. In contrast, deletion of Srf in developing B cells did not interfere with the growth and survival of B cells in general, yet led to a complete loss of marginal zone B cells and a marked reduction of the CD5+ B cell subset. Our study also revealed a contribution of SRF to the expression of the surface molecules IgM, CD19, and the chemokine receptor 4 in B lymphocytes. We conclude that SRF fulfills essential and distinct functions in the differentiation of different types of lymphocytes.

Background Although the contribution of alveolar type II epithelial cell (AEC II) activities in various aspects of respiratory immune regulation has become increasingly appreciated, our understanding of the contribution of AEC II transcriptosome in immunopathologic lung injury remains poorly understood. We have previously established a mouse model for chronic T cell-mediated pulmonary inflammation in which influenza hemagglutinin (HA) is expressed as a transgene in AEC II, in mice expressing a transgenic T cell receptor specific for a class II-restricted epitope of HA. Pulmonary inflammation in these mice occurs as a result of CD4+ T cell recognition of alveolar antigen. This model was utilized to assess the profile of inflammatory mediators expressed by alveolar epithelial target cells triggered by antigen-specific recognition in CD4+ T cell-mediated lung inflammation. Methods We established a method that allows the flow cytometric negative selection and isolation of primary AEC II of high viability and purity. Genome wide transcriptional profiling was performed on mRNA isolated from AEC II isolated from healthy mice and from mice with acute and chronic CD4+ T cell-mediated pulmonary inflammation. Results T cell-mediated inflammation was associated with expression of a broad array of cytokine and chemokine genes by AEC II cell, indicating a potential contribution of epithelial-derived chemoattractants to the inflammatory cell parenchymal infiltration. Morphologically, there was an increase in the size of activated epithelial cells, and on the molecular level, comparative transcriptome analyses of AEC II from inflamed versus normal lungs provide a detailed characterization of the specific inflammatory genes expressed in AEC II induced in the context of CD4+ T cell-mediated pneumonitis. Conclusion An important contribution of AEC II gene expression to the orchestration and regulation of interstitial pneumonitis is suggested by the panoply of inflammatory genes expressed by this cell population, and this may provide insight into the molecular pathogenesis of pulmonary inflammatory states. CD4+ T cell recognition of antigen presented by AEC II cells appears to be a potent trigger for activation of the alveolar cell inflammatory transcriptosome.

While the central role of HbA1c levels for the prediction of micro- and macrovascular complications in patients with type 1 diabetes is generally accepted; recommendations in current guidelines and the level of metabolic control actually achieved during routine care differ widely. Limited information is available on factors that influence metabolic control in the pediatric age group and during the transition from pediatric to adult diabetes care. In a large prospective multicenter database (DPV-Wiss), 338,330 individual HbA1c measurements from 27,035 patients with type-1 diabetes (94,074 observation years) were recorded between 1995 and 2005. Data were anonymously transmitted from 207 institutions. HbA1c values were mathematically standardized to the DCCT normal range (4.05–6.05%). The SAS 9.1 software was used for statistical analysis using nonparametric statistics. Median HbA1c for all measurements was 7.8%, with a strong effect of diabetes duration: median HbA1c at onset was 9.1%, during the first 2 years of diabetes 7.1% with a subsequent increase to 7.9% in patients beyond the remission phase (>2 years, 20,314 patients); a strong age dependency was present. HbA1c above the recommended guidelines was found in 23%. For all age groups, girls/women had higher HbA1c values compared to boys (mean difference 0.1%, p