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  • 1
    Electronic Resource
    Electronic Resource
    Reconstructive surgery of the lower urinary tract following exenterative resection of gynecological tumors (1999)
    Springer
    Der Urologe 38 (1999), S. 237-241 
    Details
    ISSN: 1433-0563
    Keywords: Key words Exenterative surgery • Urinary diversion • Gynecological tumors ; Schlüsselwörter Exenteration • Harnableitung • ; Gynäkologische Tumoren
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Bei fortgeschrittenen oder rezidivierenden Tumorerkrankungen der weiblichen Genitalorgane ist die pelvine exenterative Resektion dann anzustreben, wenn eine lokale Tumorinfiltration des unteren Harntraktes vorhanden ist und wenn eine Strahlentherapie mit kurativer Zielsetzung als Alternative nicht mehr zur Verfügung steht. Bei der Planung des operativen Vorgehens muß die Resektion und Rekonstruktion der betroffenen ableitenden Harnorgane mit in das gynäkologische Therapiekonzept einbezogen werden. In der vorliegenden Serie war bei 11/32 Patientinnen die Rekonstruktion der ableitenden Harnwege möglich, so daß die Harnblase mit ausreichender Funktion erhalten werden konnte. Zur Harnableitung nach vorderer Exenteration wurde bei 17/32 Patientinnen ein kontinentes Harnreservoir gewählt und bei 4/17 Patientinnen ein Ileumkonduit. Nur bei Anlage eines kontinenten Harnreservoirs war aufgrund von Komplikationen in 5 Fällen eine operative Reintervention erforderlich. Bei diesen Patientinnen bestand ein Zustand nach primärer Strahlentherapie des gynäkologischen Tumors. Bei weiteren 11 Patientinnen nach einer primären Strahlentherapie traten keine Komplikationen auf. 9 der 32 Patientinnen haben den operativen Eingriff mit einer durchschnittlichen Überlebenszeit von 40,8 (25–57) Monaten bislang tumorfrei überlebt. Kontinente Harnreservoire stellen im Rahmen einer pelvinen exenterativen Maßnahme eine exzellente Möglichkeit dar, den damit verbundenen Ausfall an Körperfunktionen zu ersetzen. Aufgrund der häufig sehr komplexen Verhältnisse v. a. infolge vorausgegangener umfangreicher therapeutischer Maßnahmen ist aber eine strenge Selektion der Patientinnen unverzichtbare Voraussetzung.
    Notes: Summary In locally advanced or recurrent tumors of the female genital tract anterior or total exenteration may be mandatory in case of tumor invasion into the lower urinary tract or if a second course of radiation therapy is not feasible. The management of resection and reconstruction of the affected lower urinary tract has to be well integrated into the gynecological therapeutic concept. In 11/32 patients the reconstruction of the partially resected lower urinary tract was feasible with preservation of a functionally intact urinary bladder. Urinary diversion following pelvic exenteration was achieved in 13/17 patients with a continent urinary reservoir and in 4/17 patients with an ileal conduit. Operative reinterventions were needed only in patients with continent urinary diversion in 5 cases. All these patients had a past history of primary radiation therapy of their gynecological tumor. In the remaining other 11 patients with a history of primary radiation therapy no complications occurred. 9 of 32 patients survived the operative procedure 40,8 (25–57) month with no evidence of recurrent tumor. Continent urinary diversion represents an excellent therapeutic option for replacement of function lost due to exenterative pelvic surgery. Stringent selection of patients is mandatory to consider the presented therapeutic concept a reasonable tool in the management of the described clinical situations.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001200050274
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  • 2
    Electronic Resource
    Electronic Resource
    Visualization of interstitial cells of Cajal in the mouse colon by vital staining (1998)
    Springer
    Cell & tissue research 292 (1998), S. 275-282 
    Details
    ISSN: 1432-0878
    Keywords: Key words Interstitial cells (Cajal) ; Large intestine ; Fluorescent dyes ; Vital staining ; Ultrastructure ; Mouse (BALB/c)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Interstitial cells of Cajal (ICCs) are believed to be a major element in generating the spontaneous rhythm of the gastrointestinal tract. A prominent problem in the study of these cells has been the difficulty in observing them in intact tissues. We used the lipophilic dye DiI to stain ICCs in the submucosal-circular muscle border of freshly dissected mouse colon. The placement of small DiI crystals in this area resulted in the labeling of ICC-like cells. Two main morphological cell types, viz., bipolar and multipolar, were noted. Bipolar cells had two primary processes emerging from the poles of an elongated soma. The mean length of these processes was 78.7 μm. These cells constituted 42.3% of the sample (n=105). Multipolar cells (54.3% of total) had a less elongated soma and extended 3–6 main processes whose mean length was 56.3 μm. These processes showed no preferred direction. The length of the primary processes of bipolar cells was 40% greater than that of multipolar cells (P〈0.02). Three cells (2.9%) had only one primary process. The DiI stain could be converted into a stable electron-opaque product. Electron-microscopic observations showed that these cells had the typical appearance of ICCs reported in previous studies. This staining method should be useful for physiological investigations of ICCs in gastrointestinal tissues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410051058
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Morphology of diaphragm neuromuscular junctions on different fibre types (1996)
    Springer
    Journal of neurocytology 25 (1996), S. 88-100 
    Details
    ISSN: 1573-7381
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We hypothesize that the morphology of the neuromuscular junction on different muscle fibre types varies, reflecting differences in activation history. In the rat diaphragm muscle, we used a three-colour fluorescent immunocytochemical technique to simultaneously visualize (1) innervating axons and presynaptic nerve terminals, (2) motor endplates and (3) myosin heavy chain isoform expression (muscle fibre type). Laser-scanning confocal microscopy was then used to optically section the triple-labelled muscle fibres, and create three-dimensional views of the neuromuscular junction. Type I fibres were innervated by the smallest axons, and type IIa, IIx and IIb fibres by progressively larger axons. Absolute planar areas of nerve terminals and endplates progressively increased from type I, IIa, IIx to IIb fibres. When normalized for fibre diameter planar areas of nerve terminals were largest on type I fibres, with no difference among type II fibres. The normalized planar area of endplates were larger for type I and IIb fibres, compared to type IIa and IIx fibres. The three-dimensional surface area of endplates was largest on type I fibres, with no differences across type II fibres. When normalized for fibre diameter, endplate surface areas increase progressively from type I, IIa, IIx to IIb fibres. The branches increased progressively from type I, IIa, IIx to IIb fibres. Conversely, individual branch length was longest on type I fibres, and shortest on type IIb fibres. The extent of overlap of pre- and postsynaptic elements of the neuromuscular junction decreased progressively on type I, IIa, IIx and IIb fibres. We conclude that these morphological differences at the neuromuscular function of different fibre types reflect differences in activation history and may underlie phenotypic differences in neuromuscular transmission.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02284788
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    Paper (German National Licenses)
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  • 4
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    High-efficiency genome editing via 2A-coupled co-expression of fluorescent proteins and zinc finger nucleases or CRISPR/Cas9 nickase pairs (2014)
    Oxford University Press
    Details
    Publication Date: 2014-06-03
    Description: Targeted endonucleases including zinc finger nucleases (ZFNs) and clustered regularly interspaced short palindromic repeats (CRISPRs)/Cas9 are increasingly being used for genome editing in higher species. We therefore devised a broadly applicable and versatile method for increasing editing efficiencies by these tools. Briefly, 2A peptide-coupled co-expression of fluorescent protein and nuclease was combined with fluorescence-activated cell sorting (FACS) to allow for efficient isolation of cell populations with increasingly higher nuclease expression levels, which translated into increasingly higher genome editing rates. For ZFNs, this approach, combined with delivery of donors as single-stranded oligodeoxynucleotides and nucleases as messenger ribonucleic acid, enabled high knockin efficiencies in demanding applications, including biallelic codon conversion frequencies reaching 30–70% at high transfection efficiencies and ~2% at low transfection efficiencies, simultaneous homozygous knockin mutation of two genes with ~1.5% efficiency as well as generation of cell pools with almost complete codon conversion via three consecutive targeting and FACS events. Observed off-target effects were minimal, and when occurring, our data suggest that they may be counteracted by selecting intermediate nuclease levels where off-target mutagenesis is low, but on-target mutagenesis remains relatively high. The method was also applicable to the CRISPR/Cas9 system, including CRISPR/Cas9 mutant nickase pairs, which exhibit low off-target mutagenesis compared to wild-type Cas9.
    Print ISSN: 0305-1048
    Electronic ISSN: 1362-4962
    Topics: Biology
    Published by Oxford University Press
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  • 5
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    Nuclease-mediated gene editing by homologous recombination of the human globin locus (2014)
    Oxford University Press
    Details
    Publication Date: 2014-01-28
    Description: Tal-effector nucleases (TALENs) are engineered proteins that can stimulate precise genome editing through specific DNA double-strand breaks. Sickle cell disease and β-thalassemia are common genetic disorders caused by mutations in β-globin, and we engineered a pair of highly active TALENs that induce modification of 54% of human β-globin alleles near the site of the sickle mutation. These TALENS stimulate targeted integration of therapeutic, full-length beta-globin cDNA to the endogenous β-globin locus in 19% of cells prior to selection as quantified by single molecule real-time sequencing. We also developed highly active TALENs to human -globin, a pharmacologic target in sickle cell disease therapy. Using the β-globin and -globin TALENs, we generated cell lines that express GFP under the control of the endogenous β-globin promoter and tdTomato under the control of the endogenous -globin promoter. With these fluorescent reporter cell lines, we screened a library of small molecule compounds for their differential effect on the transcriptional activity of the endogenous β- and -globin genes and identified several that preferentially upregulate -globin expression.
    Print ISSN: 0305-1048
    Electronic ISSN: 1362-4962
    Topics: Biology
    Published by Oxford University Press
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