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  • 1
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    German Medical Science; Düsseldorf, Köln
    In:  54. Jahrestagung der Norddeutschen Orthopädenvereinigung e.V.; 20050616-20050618; Hamburg; DOC05novP11 /20050613/
    Publication Date: 2005-06-14
    Keywords: total knee revision surgery ; modular implant system ; retrospective study ; ddc: 610
    Language: English
    Type: conferenceObject
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  • 2
    Abstract: Human papillomaviruses (HPVs) of the genus Betapapillomavirus appear to be involved in the early stages of skin cancer development, since both the prevalence and viral load are higher in precancerous actinic keratoses than in skin cancers. Interleukin-8 (IL-8) is an inflammatory cytokine that serves to alert the surrounding tissue after UV-induced damage. We examined the effects of the E2, E6 and E7 proteins of HPV8 and the E6 proteins of various HPV genotypes on IL-8 secretion from primary keratinocytes. HPV5 and HPV8 E6 showed the highest downregulation of basal IL-8 secretion. HPV8 E6 also negatively modulated IL-8 mRNA expression and protein secretion upon UVB irradiation. The downregulation of IL-8 in actinic keratoses may weaken the response to UV-induced damage and thus favour the accumulation of UVB-induced mutations.
    Type of Publication: Journal article published
    PubMed ID: 20007354
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  • 3
    Abstract: BACKGROUND: A role for cutaneous human beta-papillomavirus (HPV) types as co-factors in the development of non-melanoma skin cancer has been postulated. Here we have investigated the effects of E7 expression on keratinocyte differentiation, proliferation and cell-cycle proteins in organotypic skin cultures. METHODS: Recombinant retroviruses containing the E7 genes from cutaneous HPV types 1, 4, 5, 8, 20, 38 and RTRX7 were produced that include types associated with benign and malignant lesions. Adult human primary keratinocytes were transduced with these recombinant retroviruses and differentiated into skin-equivalents using de-epidermalised human dermis. RESULTS: Expression patterns of the basal keratinocyte marker cytokeratin 14 (CK14) were not altered by any of the viral E7 types analysed. However, expression of the early and late differentiation markers CK10 and involucrin were markedly altered in HPV 1, 4 and 38 cultures. The highest proliferation rates in basal cell layers, as judged by BrdU and Ki67 staining, were observed in HPV 1, 4 and 38 cultures. Interestingly, co-expression of cyclin E and p16(INK4a) within the same cell of the suprabasal cell layers was observed only in cultures generated using E7 of HPV 5 or HPV 8. CONCLUSION: HPV types associated with either benign or malignant lesions perturb keratinocyte proliferation and differentiation in different ways. Moreover, expression of E7 from HPV 5 or HPV 8 seem able to overcome p16(INK4a) induced cell cycle arrest in a subset of keratinocytes.
    Type of Publication: Journal article published
    PubMed ID: 19478389
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  • 4
    Keywords: RECEPTOR ; CELLS ; IN-VITRO ; SKIN ; PHOSPHORYLATION ; IDENTIFICATION ; MORPHOGENESIS ; HEPATOCYTE GROWTH-FACTOR ; C-MET ; REEPITHELIALIZATION
    Abstract: Cutaneous regeneration utilizes paracrine feedback mechanisms to fine-tune the regulation of epidermal keratinocyte proliferation and migration. However, it is unknown how fibroblast-derived hepatocyte growth factor (HGF) affects these mutually exclusive processes in distinct cell populations. We here show that HGF stimulates the expression and phosphorylation of the microtubule-destabilizing factor stathmin in primary human keratinocytes. Quantitative single cell- and cell population-based analyses revealed that basal stathmin levels are important for the migratory ability of keratinocytes in vitro; however, its expression is moderately induced in the migration tongue of mouse skin or organotypic multi-layered keratinocyte 3D cultures after full-thickness wounding. In contrast, clearly elevated stathmin expression is detectable in hyperproliferative epidermal areas. In vitro, stathmin silencing significantly reduced keratinocyte proliferation. Automated quantitative and time-resolved analyses in organotypic cocultures demonstrated a high correlation between Stathmin/phospho-Stathmin and Ki67 positivity in epidermal regions with proliferative activity. Thus, activation of stathmin may stimulate keratinocyte proliferation, while basal stathmin levels are sufficient for keratinocyte migration during cutaneous regeneration.
    Type of Publication: Journal article published
    PubMed ID: 24066165
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  • 5
    Keywords: EXPRESSION ; carcinoma ; T-CELLS ; IN-SITU ; TISSUE MICROARRAYS ; TECHNOLOGY
    Abstract: OBJECTIVE: To analyze intratumoral heterogeneity of immune cells and the resulting impact of heterogeneity on the level of individual patient prediction. STUDY DESIGN: Using whole slide imaging by virtual microscopy, we present the first spatial quantitative study of immune cells in a set of colorectal cancer primary tumors. We generated "tumor maps" based on cell densities in fields of 1 mm2, visualizing intratumoral heterogeneity. In this example, cutoffs of marker-based cell stains identified by tissue microarray (TMA) led to ambiguous decisions in 11 of the 20 patients studied. Classic TMA analysis can be used in large patient cohorts to generate clinically significant predictors. The transfer of these predictors from large-scale TMA to individualized predictions thus far has not been investigated. In colorectal cancer, TMA-based quantitative immune cell counts using immune cell surface molecules (CD3, CD8, Granzyme B, and CD45RO) have been shown to be potentially better predictors for patient survival than the classical TNM system. RESULTS: Our results make clear that for individualized prognostic evaluations, whole slide imaging by virtual microscopy is irreplaceable during identification of prognostic markers as well as in their subsequent application. CONCLUSION: In the future, spatial marker signatures could contribute to individual patient classifiers.
    Type of Publication: Journal article published
    PubMed ID: 21456345
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  • 6
    Keywords: CANCER ; EXPRESSION ; CELL ; Germany ; IN-VIVO ; THERAPY ; VIVO ; SYSTEM ; TOOL ; liver ; GENE ; GENE-EXPRESSION ; GENES ; PROTEIN ; MICE ; MACROPHAGES ; INDUCTION ; tumour ; BIOLOGY ; culture ; MOUSE ; gene expression ; VECTORS ; PROMOTER ; REGION ; VACCINES ; REGIONS ; DELIVERY ; Jun ; CARRIERS ; GREEN FLUORESCENT PROTEIN ; LUCIFERASE ; CYTOSINE DEAMINASE ; RE ; THERAPIES ; REPORTER GENE ; CARRIER ; HISTOLOGY ; in vivo ; E ; TOOLS ; spleen ; microbiology ; ENGLAND ; host ; FLUORESCENT PROTEIN ; FLUORESCENT ; PLASMIDS ; bacterial ; ARABAD PROMOTER ; tumour therapy
    Abstract: We have used Salmonella enterica serovar Typhimurium (S. typhimurium) which are able to colonize tumours besides spleen and liver. Bacteria were equipped with constructs encoding green fluorescent protein or luciferase as reporters under control of the promoter P-BAD that is inducible with L-arabinose. Reporter genes could be induced in culture but also when the bacteria resided within the mouse macrophages J774A.1. More important, strong expression of reporters by the bacteria could be detected in mice after administration of L-arabinose. This was especially pronounced in bacteria colonizing tumours. Histology demonstrated that the bacteria had accumulated in and close to necrotic areas of tumours. Bacterial gene induction was observed in both regions. P-BAD is tightly controlled also in vivo because gene E of bacteriophage Phi X174 could be introduced as inducible suicide gene. The possibility to deliberately induce genes in bacterial carriers within the host should render them extremely powerful tools for tumour therapy
    Type of Publication: Journal article published
    PubMed ID: 17298393
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  • 7
    Keywords: CANCER ; EXPRESSION ; tumor ; Germany ; IN-VIVO ; THERAPY ; VIVO ; SYSTEMS ; GENE ; GENE-EXPRESSION ; MOLECULES ; TISSUE ; MICE ; INDUCTION ; gene expression ; ESCHERICHIA-COLI ; VECTOR ; PROMOTER ; NETHERLANDS ; BIOLUMINESCENCE ; ENGINEERED BACTERIA ; Gall bladder ; In vivo imaging ; Inducible promoter ; Organ colonization ; Tumor targeted bacteria
    Abstract: The probiotic bacterium Escherichia coli Nissle 1917 (EcN) constitutes a prospective vector for delivering heterologous therapeutic molecules to treat several human disorders. To add versatility to this carrier system, bacteria should be equipped with expression modules that can be regulated deliberately in a temporal and quantitative manner. This approach is called in vivo remote control (IVRC) of bacterial vectors. Here, we have evaluated promoters P-araBAD, P-rhaBAD and P-tet, which can be induced with L-arabinose, L-rhamnose or anhydrotetracycline, respectively. EcN harboring promoter constructs with luciferase as reporter gene were administered either orally to healthy mice or intravenously to tumor bearing animals. Subsequent to bacterial colonization of tissues, inducer substances were administered via the oral or systemic route. By use of in vivo bioluminescence imaging, the time course of reporter gene expression was analyzed. Each promoter displayed a specific ill vivo induction profile depending on the niche of bacterial residence and the route of inducer administration. Importantly, we also observed colonization of gall bladders of mice when EcN was administered systemically at high doses. Bacteria in this anatomical compartment remained accessible to remote control of bacterial gene expression.
    Type of Publication: Journal article published
    PubMed ID: 19665575
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  • 8
  • 9
    ISSN: 1432-055X
    Keywords: Schlüsselwörter Sevofluran ; Inhalationseinleitung ; Schadstoffbelastung ; Kinderbronchoskopie ; Key words Sevoflurane ; Inhalation induction ; Occupational exposure ; Paediatric bronchoscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Abstract General anaesthetic agents are frequently used for paediatric bronchoscopy. A disadvantage of open-system anaesthesia is the contamination of the working environment. The aim of this study was to determine the exposure of the anaesthesiologist and endoscopist during paediatric bronchoscopy under general anaesthesia in different working environments and to compare these measurements to the currently valid international threshold limits. Materials and methods: Twenty-five children (ASA I–III) scheduled for diagnostic bronchoscopy were included in the study. After inhalational induction, all patients were intubated with a nonflexible bronchoscope and manually ventilated through a side-arm of the bronchoscope. Maintenance of anaesthesia was achieved with sevoflurane (2–3 vol.%) in pure oxygen. Trace concentrations were measured every 90 s in the breathing zones of the operating theatre (OT) personnel by means of a highly sensitive direct-reading instrument (Brüel & Kjaer 1302). The lower detection limit was 0.02 ppm. The investigation was done in an OT with and without air-conditioning and a scavenging system. Results: The mean age of the children was 50.3 months (range: 3–109 months). Ventilation and oxygenation were stable throughout the bronchoscopic procedure. Mean exposure to sevoflurane in the OT without air-conditioning and a scavenging system was over 40 ppm for the anaesthetist and 50 ppm for the endoscopist. All international threshold limit values were exceeded. Peak concentrations higher than 100 ppm could be detected during 40% of the anaesthetics. Conclusion: the main finding of the present study is that under inhalation anaesthesia with sevoflurane for paediatric bronchoscopy, occupational exposure is higher than all known health regulation guidelines permit. Therefore, the use of total intravenous anaesthesia is advocated even in very small infants.
    Notes: Zusammenfassung Die Bronchoskopie bei Kindern mittels volatiler Anästhetika ist ein gängiges und anerkanntes Verfahren. Nachteilig ist allerdings eine Kontamination der Umgebung mit diesen Substanzen. Zielsetzung: Ziel der Untersuchung war es, die Exposition des Personals sowohl in dem vorhandenen Untersuchungsraum in der Pädiatrie als auch unter optimalen lüftungstechnischen Bedingungen im OP Saal hinsichtlich der derzeit gültigen Arbeitsschutzgrenzwerte zu messen. Methodik: 25 Kinder, die für eine diagnostische Bronchoskopie vorgesehen waren, wurden in die Studie eingeschlossen. Nach inhalativer Einleitung wurden die Kinder mit einem starren Bronchoskop intubiert und über eine Bypassöffnung manuell ventiliert. Für die anschließende Gastroskopie wurden die Kinder mit nicht blockbaren Tuben intubiert. Die Narkose wurde mit Sevofluran aufrechterhalten. Die Spurenkonzentrationen wurden mittels eines direktanzeigenden Infrarotspektrometers (Brüel & Kjaer 1302) in den Atemzonen von Pädiater und Anästhesisten gemessen. Die untere Nachweisgrenze betrug für Sevofluran 0,02 ppm, die gemessenen Konzentrationen lagen im Meßbereich des Infrarotspektrometers. Ergebnisse: Das mittlere Alter der Kinder betrug 50,3 Monate (Spannweite 3–109 Monate). Die durchschnittlichen Expositionswerte lagen in dem Raum ohne Klimatechnik für den Anästhesisten deutlich über 40 ppm, für den Pädiater über 50 ppm. Spitzenwerte von Sevofluran über 100 ppm wurden in 40% der Narkosen mehrfach übertroffen. Selbst unter suffizienter Klimatechnik wurden hohe Arbeitsplatzkonzentrationen gemessen. Schlußfolgerung: Arbeitsplatzbelastungen mit Sevofluran können in Räumen mit optimaler Klimatechnik deutlich abgeschwächt werden, obwohl auch dort hohe Arbeitsplatzkonzentrationen nachgewiesen werden konnten. Um ein mögliches Gesundheitsrisiko für Beschäftigte zu minimieren, sollte bei starren Bronchoskopien auf eine total intravenöse Anästhesietechnik ausgewichen werden.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-055X
    Keywords: Schlüsselwörter Bedside laboratory analyzes ; Blutgasanalyse ; Kartusche EG7+ ; Hkt/HLM ; i-STAT Analyzer ; Key words Bedside laboratory analyse ; Blood gas tension ; Cartidge EG7+ ; Hct/CPB ; i-STAT analyser
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Abstract Exact and quick measurements of basic laboratory parameters are important in selected patients in the perioperative period. Depending on the capabilities of a hospital’s central laboratory, the anaesthesiologist may only obtain such laboratory tests after unacceptable delays. This problem may be overcome by a new bedside measurement device that has become available from i-STAT Corporation, Princeton, USA. The hand-held, battery-driven analyser accepts blood specimens that are injected into a disposable cartridge (EG7+) and measures acidity, blood gas tensions, haematocrit, and electrolytes. The aim of this study was to determine the accuracy of such measurements by comparing them with measurements obtained by conventional laboratory test methods. Methods: Heparinised arterial blood specimens were collected in duplicate from 49 surgical patients. Measurements of ionised calcium (Ca), sodium (Na), potassium (K), pH, pCO2, pO2, base excess (BE), haematocrit (Hct), and haemoglobin (Hb) obtained by the i-STAT analyser were compared with measurements from the calibrated analysers ABL 615 and EML 100 (Radiometer, Copenhagen). Because the i-STAT analyser calculates the Hb concentration from a conductometrically measured Hct, 19 blood specimens were centrifuged in order to compare test results with conventionally obtained Hct and Hb values. As the Hct test sensitivity with the i-STAT changes with diluted blood due to its low albumin concentration, Hct and Hb measurements during cardio-pulmonary bypass (CPB) must be corrected by activating an analyser-implemented correction algorithm (Hct/CPB and Hb/CPB). Correlation analysis was performed between conventional measurements and i-STAT values (Ca, Na, K, Hct, pCO2, pO2), between values that the i-STAT analyser derives (Hb, HCO3, BE) and conventionally obtained results, and between normal and CPB-corrected Hct and Hb values. Accuracy was judged according to the national quality standard, whic h requires test results to lie within the 95% confidence interval of conventional tests. Results: Each blood specimen was analysed: erroneous results or technical failures did not occur. Measurement of one set of i-STAT values required 2.5 min. Correlation coefficients (r) between conventional and i-STAT results were: 0.85 for CA, 1.0 for K; 0.86 for Na; 0.99 for pH; 0.98 for pCO2; 0.99 for pO2; 0.93 for HCO3; 0.93 for BE; 0.46 for Hb values not corrected for CPB and 0.95 for CPB-corrected Hb; and 0.74 for Hct values not corrected for CPB and 0.98 for CPB-corrected Hct. The correlation coefficient for Hct between centrifuged and CPB-uncorrected i-STAT values was 0.81 and that for CPB-corrected values was 0.98. National accuracy requirements were not met for tests of: Ca (by 0.02 mmol/l); pH (by 0.01); pO2 including hyperoxic values (by 26.7 mmHg, but were met for pO2 values 〈200 mmHg); Hb (by 1.6 g/dl); Hb/CPB (by 0.8 g/dl); and Hct (by 6.5%, but were met for Hct/CPB values). All other tests fulfilled the required standards. Conclusion: This analyser is easy to use, reliable, and portable, and therefore suitable for the operating room, for analyses during emergencies, on peripheral wards, for preclinical screening, or at times when availability of lab tests is time-consuming or limited. The test accuracy for electrolytes, blood gases, and Hb is high enough to justify routine use of the i-STAT analyser in clinical practice. That the nationally required quality standards for Ca, pH, and Hb were not met is not of importance because the measured deviation was too small to have clinical relevance. When analysing diluted blood with a low Hct and low oncotic pressure, it is important to activate the analyser’s correction algorithm „CPB”, because the obtained results will then comply with the required accuracy.
    Notes: Zusammenfassung Zielsetzung und Methoden: In der chirurgischen Operationseinheit und Intensivstation der Universitätsklinik wurden bei 49 Patienten arterielle Blutproben entnommen und simultan mit dem i-STAT Analyzer (Kartusche EG7+) und einem Referenzgerät ausgewertet. Die Kartusche EG7+ ermittelt die Parameter ionisiertes Kalzium, Kalium, Natrium, pH, pCO2, pO2 und Hämatokrit. HCO3, BE, SO2 und Hb werden aus diesen Daten errechnet. Der Hkt bzw. das Hb kann mit der speziellen Anpassung (HLM) für erniedrigtes Serumprotein z.B. während kardiopulmonalem Bypass unter Herz-Lungenmaschine (HLM) korrigiert werden. Ergebnisse: Mit dem Referenzgerät ABL 615 und EML 100 wurden die Korrelationen (r) für ionisiertes Kalzium (r=0,85), Kalium (r=1,0), Natrium (r=0,86), pH (r=0,99), pCO2 (r=0,98), pO2 (r=0,99), HCO3 (r=0,93), BE (r=0,93), Hb (r=0,46), Hb/HLM (r=0,95), Hkt (r=0,81) und Hkt/HLM (r=0,98) ermittelt. Die 95% Binominalverteilung der Differenzen lagen beim Kalium, Natrium, pCO2, pO2 und Hkt/HLM innerhalb der Vorgaben zur Qualitätssicherung der Bundesärztekammer. Die Meßwerte für ionisiertes Kalzium, pH und Hb/HLM lagen zwar außerhalb der empfohlenen Toleranzen der BÄK, sind aber für den klinischen Gebrauch akzeptabel. Im anästhesiologischen Bereich sollte zur Hkt- oder Hb Bestimmung immer die „HLM” Anpassung für erniedrigtes Serumprotein programmiert werden. Zusammenfassung: Der einfache Gebrauch, die Transportierbarkeit und eine Analysezeit der Blutproben von ca. 2,5 min sowie die Wirtschaftlichkeit bei geringstem Wartungsaufwand prädestinieren dieses Gerät auch für innerklinische Notfälle, z.B. im Schockraum oder auf Normalstation.
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