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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Bundesgesundheitsblatt, Gesundheitsforschung, Gesundheitsschutz 42 (1999), S. 911-921 
    ISSN: 1437-1588
    Keywords: Schlüsselwörter ; Verwilderte Haustauben (Columba livia domestica) ; Bestandsregulierung ; Stadthygiene ; Gesundheitsschutz ; Ökologisches Management ; Key words ; Feral pigeons ; Street pigeons (Columba livia domestica) ; Population reduction ; Habitat manipulation ; Urban hygiene ; Public health ; Bird management strategies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary The report, in two parts, sums up the findings of a survey covering 52 cities in Central Europe. Most administrations have imposed a ban on feeding street pigeons, but this has been more or less unsuccessful. Information campaigns were used on a large scale, along with more controversial methods such as catching the birds or shooting them down, providing pigeon houses, administering hormone preparations, chemosterilization, and exclusion from buildings. Few attempts were made to reduce the number of breeding places. Population reduction by poisons or natural enemies seems to be negligible. The findings of the survey are compared with results reported in the specialist literature, followed by a discussion of the ecological, hygienic and legal aspects, including animal protection, pest control, and public health. The strategy proposed to improve the present situation, which is found unsatisfactory in many respects, is to organize along the principles of ecological management. By optimizing the available options and following scientific criteria, it aims to combine those methods which appear most promising under local conditions. Any drawbacks are to be compensated by the enhancement of positive elements to give the best possible overall results.
    Notes: Zusammenfassung Der zweiteilige Bericht faßt die Ergebnisse einer Umfrage zusammen, bei der sich die Verwaltungen von 52 mitteleuropäischen Städten zum Straßentaubenproblem geäußert haben. Überwiegend versucht man mehr oder minder erfolglos, eine Bestandsverminderung durch Taubenfütterungsverbot zu erreichen. Häufig wird auch von Bürgerbelehrungen und Informationskampagnen berichtet. Ferner spielen Fangaktionen, Taubenhäuser, Hormonpräparate und Chemosterilantien, Abschuß und Absperrmaßnahmen an Gebäuden eine meist ambivalente Rolle. Bei der Verminderung der Brutplatzangebote sind nur schwache Bemühungen erkennbar. Natürliche Feinde (Greifvögel) scheinen in den wenigsten Städten eine Rolle bei der Reduzierung von Straßentauben zu spielen. Vergiftungen werden offenbar nicht mehr vorgenommen. Die Erfahrungen der Kommunen werden mit den in der Fachliteratur vorliegenden Ergebnissen verglichen. Kommentare aus ökologischer und hygienezoologischer Sicht berücksichtigen die rechtlichen Grundlagen von Tierschutz, Schädlingsbekämpfung und Gesundheitsvorsorge. Um die insgesamt noch sehr unbefriedigende Situation bei der Bestandsverminderung von verwilderten Haustauben zu verbessern, wird ein Strategieprogramm nach den Organisationsprinzipien des „ökologischen Managements” vorgeschlagen. Hierzu sollten sämtliche anwendbaren Möglichkeiten nach wissenschaftlichen Kriterien optimiert und den jeweils stadtspezifischen Umständen entsprechend so miteinander kombiniert werden, daß methodische Nachteile durch die Vorteile anderer Komponenten ausgeglichen werden und das Gesamtkonzept nach dem Verstärkerprinzip Optimalergebnisse erzielt.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Bundesgesundheitsblatt, Gesundheitsforschung, Gesundheitsschutz 42 (1999), S. 902-910 
    ISSN: 1437-1588
    Keywords: Schlüsselwörter ; Trinkwasser ; Bleileitungen ; Key words ; Drinking water ; Lead pipes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary On 3.11.1998 the Drinking Water Directive passed the European Parliament. This new directive will lower the current limit value for lead in Germany from 0,040 mg/l to 0,025 mg/l in 2003 and to 0,010 mg/l in 2013. Since there are still around 7800 premises with plumbing-systems containing lead-pipes in Frankfurt, the Stadtgesundheitsamt started the „Frankfurt Lead-Project” in 1997. Aim of the project was the investigation and, if necessary, the order to exchange all concerned plumbing systems. The owners of the buildings are liable to a fee for these measures of the local public health service, thus covering of the costs should be attainable. Within the project 3 to 5 water probes from each concerned building were sampled, to take the stagnation-problem into account. The drinking water of 50% of the investigated flats reached the EG-parametric value of 0,01 mg/l, which has to be transferred into national law from 2013 on the latest. Houses with plumbing-systems not containing lead were part of this sample. A sample of houses with only lead containing plumbing systems would result in considerably higher values. The mean lead-concentration after a stagnation-period of 3 hours was 0,034 mg/l and therefore significantly exceeded the EG-parametric value of 0,025 mg/l. Experts agree that the future EG-parametric values can not be met with special water treatments, but only by replacement of lead pipes by pipes composed of other suitable materials. Even pipe-coating techniques can not be recommended at this time. To our experience the new EG-parametric-value for lead of 0,025 mg/l, which has to be transferred into national law by the european member states from 2003 on, will require the sanitation of nearly every house with a lead-containing plumbing system. All the more so because the EG-parametric-value of 0,10 mg/l will become effective from 2013 on, what factually equals a prohibition of lead-pipes. Considering the time given by the EG-Drinking Water Guideline and the experience reported here we conclude that compliance with the new EG-parametric values would not be possible for most of the German cities and communities having a lead-pipe problem. It is foreseeable therefore, that the Federal Republic of Germany will have to face complaints for non-compliance with the parametric value from the European Commission. Based on the experiences with the lead-project we would recommend the prohibition of lead pipes whithin domestic distribution systems as an appropriate measure to achieve compliance with the parametric value for lead. Such a prohibition would make it possible to meet the parametric value for lead, create a reliable legal situation for both consumers and owners of the concerned plumbing systems, save considerable expenses for staff and laboratory tests, make the exchange of lead-containing installations easy to plan and would avoid conflicts about the interpretation of monitoring results.
    Notes: Zusammenfassung Durch die am 3.11.1998 vom Europäischen Parlament verabschiedete Richtlinie des Rates der Europäischen Gemeinschaft über die Qualität von Wasser für den menschlichen Gebrauch muß eine Absenkung des Bleigrenzwertes in Deutschland von derzeit 0,040 mg/l auf 0,025 mg/l bis spätestens zum Jahr 2003 und auf 0,010 mg/l bis spätestens zum Jahr 2013 erfolgen. Da in Frankfurt a.M. bekannt war, daß noch ca. 7800 Liegenschaften mit bleihaltigen Hausinstallationen ausgestattet sind, wurde ab 1997 im Stadtgesundheitsamt das „Blei-Projekt” gestartet, mit dem eine Untersuchung und ggf. erforderliche Sanierung aller betreffenden Häuser binnen max. zehn Jahren ab 1996 unter dem Aspekt der Kostendeckung durchgesetzt werden soll. Pro untersuchtem Haus wurden mehrere Proben entnommen, um das Stagnationsproblem adäquat zu berücksichtigen. In 50% der von uns untersuchten Wohnungen, unter denen sich auch Hausinstallationen ohne Bleileitungen befanden, wurde der ab dem Jahr 2013 geltende Grenzwert von 0,010 mg/l bereits erreicht. Bei einer nur aus bleihaltigen Hausinstallationen bestehenden Stichprobe ist aufgrund unserer Daten zu erwarten, daß dieser Wert sogar deutlich überschritten würde. Der Mittelwert der Leitungsproben nach 3 Stunden Stagnation überschreitet mit 0,034 mg/l den ab dem Jahr 2003 geltenden Grenzwert von 0,025 mg/l klar. Aus der Fachdiskussion ist bekannt, daß sich die zukünftigen Grenzwerte nicht mit Aufbereitungsmaßnahmen wie Phosphatierung u.ä. einhalten lassen werden, sondern daß der Austausch der Bleileitungen gegen Leitungen aus geeigneten Werkstoffen der einzig in Frage kommende Sanierungsweg ist. Auch mit Innenbeschichtungen arbeitende Sanierungstechniken können derzeit nicht empfohlen werden. Schon die Festsetzung des Trinkwasser-Grenzwertes für Blei von zunächst 0,025 mg/l ab dem Jahr 2003 bedeutet daher, daß nahezu jede bleihaltige Hausinstallation bis dahin durch vollständiges Austauschen der Bleirohre saniert werden muß. In jedem Fall gilt dies für die Einführung des Parameterwertes von 0,010 mg/l ab dem Jahr 2013, was faktischen einem Verbot von Bleileitungen gleichkommt. Aus den gegebenen Fristen und den hier gemachten Erfahrungen hinsichtlich der benötigten Bearbeitungszeiten ist die Erkenntnis abzuleiten, daß ein Vollzug der Trinkwasserverordnung ohne ein möglichst auf klare europaweite Rechtsvorschriften gegründetes Verbot in den meisten betroffenen Bundesländern, Kreisen und kreisfreien Städten nicht möglich sein wird und somit die seitens der die Bundesrepublik Deutschland als EG-Mitgliedsstaat eingegangenen Verpflichtung zur Einhaltung des Parameterwertes für Blei nicht erfüllt werden kann. Ein Verbot für Bleileitungen würde die Einhaltung des Parameterwertes ermöglichen, Rechtsklarheit sowohl für die Verbraucher als auch für die Eigentümer von Hausinstallationen bringen, erhebliche Mittel für Untersuchungs- und Verwaltungskosten einsparen lassen, die ohnehin unumgängliche Sanierung von bleihaltigen Hausinstallationen für die Eigentümer besser planbar machen sowie Rechtsstreitigkeiten über die Interpretation von Meßwerten vermeiden.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1436-2813
    Keywords: thyroid tumor ; apoptosis ; TUNEL ; MIB-1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To clarify the growth mechanisms of thyroid tumors, we examined apoptotic cells in 61 thyroid tumors, consiting of 14 adenomas, 35 papillary carcinomas, 4 follicular carcinomas, and 8 undifferentiated carcinomas, using terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate digoxigenin-nick end labeling (TUNEL). The proliferative activity was also evaluated immunohistochemically using the monoclonal antibody to Ki-67 antigen (MIB-1) in the same tumors. The apoptotic index (AI) was expressed as a percentage of the TUNEL-positive cells in the tumor cells, and a proliferation index (PI), being the percentage of Ki-67 positive cells, was calculated for each tumor. The overall level of AI was very low in all histotypes of the thyroid tumors analyzed, the mean AI being 0.5±0.4 in adenoma, 0.4±0.3 in differentiated carcinoma, and 1.8±1.5 in undifferentiated carcinoma. The PI in the thyroid tumor subtypes was significantly lower in adenoma and differentiated carcinoma, at 0.5 ±0.7 and 1.1±0.7, respectively, than that in undifferentiated carcinoma at 14.5±3.7 (P〈0.05). There was no correlation between clinicopathological factors and AI or PI in differentiated thyroid carcinoma. Our findings suggest that apoptosis occurs infrequently in thyroid tumors, and that proliferative activity markedly differs according to the thyroid tumor subtypes. Moreover, the ratio between proliferating cells and apoptotic cells may reflect thyroid tumor progression.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1436-2813
    Keywords: Key Words: thyroid tumor ; apoptosis ; TUNEL ; MIB-1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: P 〈 0.05). There was no correlation between clinicopathological factors and AI or PI in differentiated thyroid carcinoma. Our findings suggest that apoptosis occurs infrequently in thyroid tumors, and that proliferative activity markedly differs according to the thyroid tumor subtypes. Moreover, the ratio between proliferating cells and apoptotic cells may reflect thyroid tumor progression.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Arthroskopie 12 (1999), S. 279-288 
    ISSN: 1434-3924
    Keywords: Schlüsselwörter ; Knorpelschaden ; Knie ; Knorpelregeneration ; Key words ; Cartilage damage ; Knee ; Cartilage regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Damage to articular cartilage is a common problem. Damage to a joint surface can lead to premature arthritis. In this review the results of the different forms of therapy for cartilage damage such as lavage, drilling, microfracturing, abrasion arthroplasty, transplantation of osteochondral allografts, periosteal arthroplasty and autologous chondrocyte transplantation are introduced to the reader.
    Notes: Die Therapie des Knorpelschadens stellt ein allgemeines Problem dar. Beschädigte Knorpeloberflächen können zur frühzeitig auftretenden Arthrose führen. In dieser Übersichtsarbeit werden die einzelnen Therapieformen zur Behandlung von Knorpelschäden wie die Lavage, Knochenmarkstimulationstechniken (subchondrale Bohrung, Mikrofrakturierung, Abrasionsarthroplastik) sowie neuere Therapieformen wie die Transplantation autologer Knochen-Knorpel-Zylinder, die Periostlappenplastik und die autologe Chondrozytentransplantation mit bisherigen Ergebnissen vorgestellt. Der Leser soll somit einen Überblick über diese Verfahren erhalten.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1434-3924
    Keywords: Schlüsselwörter ; Rehabilitation ; Humane Fibroblasten ; Zyklische mechanische Dehnung ; Zellproliferation ; Key words ; Rehabilitation ; Human fibroblast ; Cyclic strain ; Cell proliferation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Experimental and clinical studies document the benefits of early controlled motion and loading in the treatment of musculoskeletal injuries. At the cellular level, dynamic strain plays a key role in cell stimulation and organization of the extracellular matrix. Although the positive effects of physical strain on tendon tissue are well known, little knowledge exists as to how mechanical strain affects tendon cells. The aim of the present study was to investigate in vitro the influence of cyclic strain on cell proliferation of human tendon fibroblasts. Human tendon fibroblasts from patellar tendon were cultured on silicone dishes. Cyclic biaxial strain was applied to the dishes in their long axis with third passage cells. A 5% strain magnitude and a frequency of 1 Hz were applied. The time of applied strain varied between 15, 30 and 60 min. After the fibroblasts were put under strain, the cells were tested for proliferation after 6, 12 and 24 h using incorporation of 5-bromo-2′-deoxy-uridine. As a control, cells were grown on silicone dishes but did not receive any strain. A biphasic response in proliferation was observed for the 15- and 60-min strain periods. At 6 and 24 h the proliferation was increased compared to the controls. The proliferation was decreased at 12 h. After a strain duration of 30 min the proliferation was inferior to that of the controls at all times measured. Application of mechanical stress to tendon fibroblasts results in an alteration of cellular proliferation, depending on the stress time. This study will contribute to an understanding of the cellular mechanism underlying the mechanical stimulation of tendon and ligament healing.
    Notes: Klinische und tierexperimentelle Studien haben den positiven Einfluß einer frühfunktionellen Behandlung nach Verletzungen des Binde- und Stützgewebes gezeigt. Dehnung ist dabei der zentrale Stimulus für reaktive Prozesse auf zellulärer Ebene. Zyklische mechanische Dehnung führt zu einer Reihe von Reaktionen, die für die Adaption von Zellen und Geweben an unterschiedliche Belastungen und für den Heilungsprozeß von Bedeutung sind. Ungeklärt ist jedoch, wieviel Dehnungsstreß notwendig ist. Ziel dieser Studie war, den Einfluß der Dauer von definierter zyklischer Dehnung auf die Zellproliferation von humanen Fibroblasten zu untersuchen. Fibroblasten wurden dazu aus Patellarsehnen isoliert und auf Silikonschalen kultiviert. Die Silikonschalen mit Zellen der 3. Passage wurden mit einem elektromechanischen Stimulationsgerät zyklisch mechanisch in der Längsachse gedehnt. Eine Dehnungsamplitude von 5% und die Frequenz 1 Hz wurden für alle Experimente gewählt. Die Dauer der zyklischen Dehnung wurde mit 15, 30 und ¶60 min verändert. Nach insgesamt 6, 12 und 24 h wurde der Versuch beendet. Zur Quantifizierung der Zellproliferation wurde die DNA-Synthese, d. h. der ¶Einbau von 5-Bromo-2′-deoxy uridine¶(BrdU) in die DNA gemessen. Als Kontrolle dienten humane Fibroblasten auf Silikonschalen ohne mechanische Dehnung. Nach 15 und 60 min zyklischer Dehnung zeigte sich ein biphasischer Verlauf hinsichtlich der Zellproliferation. Nach 6 und 24 h war eine Zunahme der Zellproliferation im Vergleich zu 12 h vorhanden. 30 min zyklische mechanische Dehnung hatte im Vergleich zur Kontrolle dagegen keinen positiven Einfluß auf die Zellproliferation. Zyklische mechanische Dehnung führt in Abhängigkeit von der Streßdauer zu Veränderungen bei der Zellproliferation. Die längere Streßdauer induziert potentiell Reaktionen, die eine protektive Wirkung für die Zellproliferation haben. Die zelluläre Streßantwort basiert jedoch auf komplexen Prozessen, deren Regulation und Modulation noch nicht geklärt sind. Das Verständnis der zellulären Reaktionen auf mechanische Dehnung ist von grundlegender Bedeutung für eine funktionelle Behandlung von Sehnen- und Bandverletzungen.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Arthroskopie 12 (1999), S. 294-298 
    ISSN: 1434-3924
    Keywords: Schlüsselwörter ; Knie ; Knorpelläsion ; Arthroskopie ; Knorpeltransplantation ; Key words ; Knee ; Cartilage lesion ; Arthroscopy ; Cartilage transplantation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: The clinical application of cartilage repair procedures is currently being discussion. To define cartilage lesions with respect to localization, degree of cartilage damage, size, and etiology we retrospectively reviewed data from 4408 patients who had knee arthroscopies. Chondral lesions were divided into traumatic (n = 572, mean age 30.2 years; 13.2–52.7 years) and degenerative (n = 976, mean age 51.4 years; 27.1–79.9 years) lesions. All traumatic cartilage lesions were associated with isolated anterior cruciuate tear. The time interval between injury and arthroscopy was subdivided in patients with acute ¶anterior cruciate ligament tear (〈 6 weeks) or with chronic joint instability (〉 6 weeks). Isolated degenerative cartilage lesions were subdivided in patients aged ≤ 40 years and 〉 40 years. Patients with other associated lesions were excluded. Because we believe that patients with grade III cartilage lesions under the age of 40 years represent the ideal type of patient to receive cartilage or osteochondral transplantation, the data set was analyzed further to characterize these patients. In both groups (traumatic and degenerative) the medial femoral condyle and the medial tibial plateau were most affected. Grade III lesions increased in number and size in all compartments with age by about 2–3.5-fold. The medial tibial plateau showed a significantly higher alteration of about 13-fold in patients aged 〉 40 years. The two subgroups comprising acute and chronic anterior cruciate ligament tears did not demonstrate any statistically significant differences in any parameter. Potential candidates for cartilage grafting accounted for 82 patients (5.3%) on the medial femoral condyle and for 33 patients (2.1%) on the medial tibial plateau of all cartilage lesions (n = 1548 in this data set. In conclusion, we found no relationship between time of surgery and degree of traumatic cartilage lesions after anterior cruciate ligament tear. The development of grade III cartilage lesions in patients older than ¶40 years was significantly higher on the medial tibia plateau than in the other compartments. That means that extrinsic factors have more influence on the cartilage damage than genetically determined factors. In respect to the observation that most of the patients achieve a total knee arthroplasty because of osteoarthritis on the medial compartment, all cartilage reconstructive procedures should be focused on the medial compartment.
    Notes: Die Indikation zur Knorpeltransplantation bzw. zum Einsatz anderer biologisch rekonstruktiver Maßnahmen wird kontrovers diskutiert. Zur Bestimmung des Musters der Knorpelschädigungen im Kniegelenk war es Ziel dieser Studie, die Parameter Lokalisation, Schädigungsgrad und Größe in Abhängigkeit von der Ätiologie zu bestimmen. Es wurden 4408 Patienten mit Kniegelenkarthroskopien ausgewertet, bei denen bei 572 Patienten (Durchschnittsalter 30,2 Jahre; 13.2–¶52,7 Jahre) eine traumatische und bei 976 Patienten (Durchschnittsalter 51,4 Jahre; 27,1–79,9 Jahre) eine degenerative Knorpelschädigung vorlag. Den traumatische Knorpelläsionen wurden Patienten mit einem gesicherten Rotationsereignis und einer isolierten assoziierten vorderen Kreuzbandläsion zugeordnet. Diese Gruppe wurde entsprechend dem Zeitintervall zwischen Ereignis und Arthroskopie in akute (〈 6 Wochen) und chronische (〉 6 Wochen) Zustände unterteilt. Den degenerativen Knorpelläsionen wurden Patienten mit einer isolierten Knorpelpathologie zugeordnet und nach dem Lebensalter ¶(≤ 40 Jahre oder 〉 40 Jahre) weiter differenziert. Patienten mit einer zusätzlichen pathologischen Veränderung wurden ausgeschlossen. Sowohl bei traumatischen als auch degenerativen Erkrankungen waren der mediale Femurkondylus und das mediale Tibiaplateau am häufigsten alteriert. Bei der altersabhängigen Untersuchung zeigte sich, daß erwartungsgemäß in allen Kompartmenten die Häufigkeit von Grad-III-Läsionen um den Faktor 2–3,5 zunahm, auf dem medialen Tibiaplateau jedoch um den Faktor 13. Der Vergleich von akuten mit längerzeitig bestehenden traumatischen Knorpelverletzungen erbrachte keinen statistisch signifikanten Unterschied in allen untersuchten Parametern. Insgesamt hatten nur 82 Patienten (5,3%) am medialen Femurkondylus und 33 Patienten (2,1%) am medialen Tibiaplateau eine Grad-III-Läsion, welche potentiell von allen Knorpelschädigungen (n = 1548) für eine Knorpeltransplantation in Frage kommen würden. Zusammenfassend ist zu schlußfolgern, daß ein Einfluß der Zeitdauer nach traumatischer Knorpelläsion mit assoziierter vorderer Kreuzbandläsion auf die untersuchten Parameter in dieser retrospektiven Untersuchung nicht nachgewiesen werden konnte. Die Entwicklung von degenerativ bedingten Grad-III-Läsionen nimmt am medialen Tibiaplateau im Vergleich zu den anderen Kompartmenten nach dem 40. Lebensjahr sprunghaft zu, so daß der Einfluß exogener Faktoren wahrscheinlicher ist als das alleinige Vorliegen einer anlagebedingten Minderbelastbarkeit des Knorpels. Im Zusammenhang mit der Beobachtung, daß die meisten Patienten aufgrund einer Varusgonarthrose endoprothetisch versorgt werden, sollte die Indikation für knorpelrekonstruierende Maßnahmen auf das mediale Kompartment fokusiert werden.
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  • 8
    ISSN: 1434-3924
    Keywords: Schlüsselwörter ; Discus-articularis-Verlagerungen ; Arthroskopie ; Lysis & Lavage ; Key words ; Discus articularis ; Displacement ; Arthroscopy ; Lysis & Lavage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Displacement of the discus articularis with and without repositioning represents a common pathological alteration of the temporomandibular joint (tmj). The primary clinical symptoms are pain and reduced jaw movement. Particularly in cases of long-term complaints, conservative treatment methods such as splint and physiotherapy often do not produce the desired results. For these patients, arthroscopy combined with lysis and lavage is the treatment of choice for diagnostic as well as therapeutic purposes. In our department we performed this surgery on a total of 23 patients with discus displacement. With this low-risk procedure we achieved a definite improvement in symptoms in 20 patients; 11 of those patients were eventually complaint-free. Additionally, we can report a general improvement in opening the mouth. Except for a slight postoperative soft-tissue swelling, no complications developed. Thus, we can conclude that arthroscopic lysis and lavage, a low-risk and minimally invasive procedure, should be preferred to open arthrotomy in the primary treatment of tmj-displacements.
    Notes: Verlagerungen des Discus articularis mit und ohne Reposition stellen eine oft zu beobachtende pathologische Veränderung des Kiefergelenks dar. Klinisch stehen dabei Schmerzhaftigkeit und Einschränkungen der Unterkieferbewegungen im Vordergrund. Häufig können diese Beschwerden gerade bei längerem Bestehen durch konservative Behandlungsmaßnahmen wie eine Schienen- oder Physiotherapie nur geringfügig beeinflußt werden. Die Arthroskopie eröffnet hier in Kombination mit einer Lysis und Lavage sowohl diagnostische als auch therapeutische Möglichkeiten. An unserer Klinik wurden insgesamt 23 Patienten mit einer Diskusluxation arthroskopiert. Bei 20 Patienten konnte mit diesem risikoarmen Eingriff eine deutliche Besserung der Symptomatik, bei 11 davon sogar eine subjektive Beschwerdefreiheit erzielt werden. Auch funktionell konnte z. B. eine generelle Verbesserung der Mundöffnung erzielt werden. Abgesehen von geringfügigen postoperativen Weichteilschwellungen waren keine Komplikationen zu verzeichnen. Als Konsequenz unserer Ergebnisse sollten die arthroskopische Lysis und Lavage als risikoarmes, minimalinvasives Verfahren bei Diskusluxationen des Kiefergelenks primär einer offenen Arthrotomie vorgezogen werden.
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  • 9
    ISSN: 1434-3924
    Keywords: Schlüsselwörter ; Radiusfraktur loco typico ; Skapholunäre Bandverletzungen ; Arthroskopisch assistierte Versorgung ; Mittelfristige Ergebnisse ; Key words ; Distal radius fracture ; Lesions of the scapholunate ligaments ; Wrist arthroscopy ; Midterm results
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Since 1993, 30 patients with acute intraarticular distal radius fractures have been treated under arthroscopic assistance. In 12 patients (40%) concomitant lesions of the intrinsic scapholunate ligaments were diagnosed. According to Geissler we found grade I-tear in one patient, grade II in three, grade III in six and grade IV in two patients. The eight individuals with grade III-and IV-lesions additionally revealed marked instablity intraoperatively. Therefore, operative stabilization was subsequently performed by temporary scapholunate and scaphocapitate arthrodesis. Seven patients out of this group (87.5%) were followed up clinically and radiologically 3 years postoperatively, on average. Clinical examination included range of motion tests and a subjective questionnaire concerning pain and working ability. Objective grip strength was measured using a Jamar tester and compared to the contralateral wrist. Radiological evaluation consisted of the posteroanterior and lateral view as well as stress views in radial and ulnar deviation. Data were evaluated by the Jakim and Cooney scoring system as well as the demerit point system by Gartland and Werley as modified by Sarmiento et al. According to Gartland, 100% of our patients showed an excellent result; these rates were 86% using the Jakim score and 60% using the Cooney scoring system. According to the subjective questionnaire all of the patients revealed an excellent or good result.
    Notes: Seit 1993 wurden an unserer Klinik 30 Patienten mit frischen distalen intraartikulären Radiusfrakturen arthroskopisch assistiert versorgt. Begleitende Verletzungen der skapholunären intrinsischen Bandverbindungen fanden sich bei 12 Patienten (40%). Entsprechend der Einteilung von Geissler fanden wir einmal eine Läsion vom Grad I, 3mal Grad II, 6mal Grad III und 2mal Grad IV. Die 8 Patienten mit den dritt- bzw. viertgradigen Läsionen zeigten intraoperativ zusätzlich eine deutliche Instabilität im Skapholunargelenk (SL-Gelenk), so daß eine temporäre skapholunäre bzw. skaphokapitäre Spickdrahtarthrodese in derselben Sitzung durchgeführt wurden. Aus dieser Gruppe konnten 7 Patienten (87,5%) nach durchschnittlich 3 Jahren klinisch und radiologisch nachuntersucht werden. Die klinische Untersuchung umfaßte ¶den Bewegungsumfang, subjektive Schmerzangaben sowie eine Befragung nach der Beschäftigung. Mittels Jamar-Dynamometer (Cedaron ® ) wurde die objektive Griffstärke beim Faustschluß im Vergleich beider Hände ermittelt. Zusätzlich wurden Röntgenbilder im a.-p.- und seitlichen Strahlengang sowie Streßaufnahmen in Radial- und Ulnarduktion angefertigt. Die erhobenen Daten wurden mittels der Scores von Jakim , Cooney sowie dem nach Sarmiento modifizierten Gartland-Werley-Score evaluiert. Nach Jakim zeigten 86% der Patienten ein sehr gutes oder gutes Resultat, nach Gartland 100% und nach Cooney 60% ein sehr gutes Ergebnis. In der subjektiven Befragung zeigten alle Patienten ein exzellentes oder gutes Ergebnis.
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  • 10
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    Journal of neural transmission 106 (1999), S. 1-21 
    ISSN: 1435-1463
    Keywords: Keywords: Paraquat ; Parkinson's disease ; transcription factor ; AP-1 ; apoptosis ; cycloheximide ; genistein ; SOD ; catalase ; oxidative stress.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary. Drugs and certain environmental toxins may be responsible for the pathogenesis of Parkinson's disease. We have used paraquat as a model toxin for this study since paraquat has been shown to make its way to the nerve terminals and cause cell death of dopamine neurons by oxidative injury. We have shown by the electrophoretic mobility shift assay that paraquat, together with low concentrations of chelated iron (Fe++/DETAPAC), induced the activation of transcription factor AP-1 binding activity to DNA. Under similar conditions we also found by both a DNA laddering assay procedure and by terminal deoxynucleotidyl transferase assay (TUNEL assay) that paraquat also induces apoptotic cell death. Interestingly, both apoptotic cell death and AP-1/DNA binding activity induced by paraquat were blocked by cyclohexamide and genistein, indicating that both the AP-1/DNA binding activation and apoptosis induced by paraquat are closely related. Moreover, cells were also protected from paraquat toxicity in the presence of antioxidant defense enzymes SOD and catalase. The results support the hypothesis that oxidative stress may be contributing to the apoptotic cell death of dopaminergic neurons, leading to the manifestation of Parkinson's disease. Since paraquat was an important herbicide in the mid 20th Century, our results have the important implication that exposure to environmental toxins such as paraquat may induce Parkinson's disease.
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  • 11
    ISSN: 1572-8773
    Keywords: cadmium ; apoptosis ; RT-PCR ; p53 gene expression ; testes ; rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Reverse transcription (RT) PCR technique was used to investigate the mechanism of apoptosis induced by Cd and the change of its related genes in testes and prostate of rats. Adult male rats were given a single (s.c.) injection of CdC l2 0, 2.5, 5.0, 10 μmol/kg. 48 h and 72 h after administration of Cd, animals were sacrificed. The results indicated that Cd can induce apoptosis in testes via p53-independent pathway. No apoptosis occurred in prostate in any of the Cd-exposed groups. There was a clearly negative relationship in testes between p53 gene expression and Cd exposure and this dose-response relationship was observed both at 48 h and 72 h. There was a very small increase of this gene expression in the dorsolateral lobe of the prostate in Cd exposed groups. The other apoptosis related gene, bcl-x, was not detectable in either control or Cd-exposed group in testes and dorsal prostate. Although the MT-I gene was expressed in testes or dorsal prostate both in control and exposed groups, no overexpression of MT-I gene was found after administration of Cd . The expression of MT-I in the ventral prostate was not detected in the control group, but a weak expression was found after Cd exposure. Since p53 is a tumo r suppressor gene which can inhibit tumorigenesis, the consequence of a Cd-induced decrease of p53 in testes may have a relation to the known risk of Cd tumorigenesis in this tissue.
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  • 12
    ISSN: 1569-8041
    Keywords: AML ; apoptosis ; etoposide ; γ-GCS ; glutathione ; oxidative stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Etoposide mediates its cytotoxicity by inducing apoptosis. Thus, mechanisms which regulate apoptosis should also affect drug resistance. Oxidants and antioxidants have been shown to participate in the regulation of apoptosis. We were interested in studying whether responsiveness of acute myeloblastic leukemia (AML) cells to etoposide is mediated by oxidative stress and glutathione levels. Patients and methods: Two subclones of the OCI/AML-2 cell line which are etoposide-sensitive (ES), and etoposide-resistant (ER), were established by the authors at the University of Oulu, and used as models. Assays for apoptosis included externalization of phosphatidylserine (as evidenced by annexin V binding), and caspase activation as indicated by cleavage of poly(ADP-ribose)polymerase (Western blotting). Peroxide formation was analyzed by flow cytometry. Glutathione and gamma-glutamylcysteine synthetase (γ-GCS) levels were determined spectrophotometrically and by Western blotting, respectively. Results: Etoposide-induced apoptosis was evident 12 hours after treatment in the ES subclone, but was apparent in the ER subclone only after 24 hours. The basal glutathione and γ-GCS levels were higher in the ER than the ES subclone. Etoposide increased peroxide formation in both subclones after 12-hour exposure. Significant depletion of glutathione was observed in the ES subclone during etoposide exposure, while glutathione levels were maintained in the ER subclone. In neither of the subclones was induction of γ-GCS observed during 24-hour exposure to etoposide. Furthermore, the catalytic subunit of γ-GCS was cleaved during apoptosis, concurrent with depletion of intracellular glutathione. When glutathione was depleted by treatment with buthionine sulfoximine, a direct inhibitor of γ-GCS, the sensitivity to etoposide was increased, particularly in the ER subclone. Conclusions: The results underline the significance of glutathione biosynthesis in the responsiveness of AML cells to etoposide. The molecular mechanisms mediating glutathione depletion during etoposide exposure might include the cleavage of the catalytic subunit of γ-GCS.
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  • 13
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    Annals of oncology 10 (1999), S. 495-503 
    ISSN: 1569-8041
    Keywords: antisense ; apoptosis ; bcl-2 ; lymphoma ; leukaemia ; phase I
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
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  • 14
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    Annals of oncology 10 (1999), S. 1011-1021 
    ISSN: 1569-8041
    Keywords: apoptosis ; chemosensitivity ; cytotoxicity ; p53
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Although hematologic malignancies and some solid tumors such as germ cell tumors and pediatric malignancies can be cured by cytotoxic treatment, the most prevalent solid tumors are relatively resistant to these interventions. Apoptosis is involved in the cell kill of anticancer drugs and p53 is believed to be of principal importance in this process. However p53 also plays a role in cell cycle arrest and DNA repair, cellular processes that can decrease the sensitivity to chemotherapy. Therefore, p53 may play a dual role after exposure to cytotoxic treatment, activating either mechanisms that lead to apoptosis or launching processes directing to DNA repair and survival of the cell. Design: In this article, we review in details the p53functions involved in the mediation of chemosensitivity. The preclinical and clinical data published in the recent years about the relation between p53 and chemosensitivity are discussed and the potential pitfalls associated to most of these studies, and that may account for the contradictory results produced so far are also mentioned.
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  • 15
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    Annals of oncology 10 (1999), S. 31-38 
    ISSN: 1569-8041
    Keywords: apoptosis ; lanreotide treatment ; neuroendocrine gastrointestinal tumors ; octreotide ; somatostatin analogs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Neuroendocrine gastrointestinal tumors express somatostatin receptors (ssts) in 80%–90% of cases and somatostatin analogs have become increasingly important in the management of these patients. Most of the presently available somatostatin analogs (octreotide, RC-160, and lanreotide) bind to the sst2 and sst5, and in higher doses to sst3 of the ssts 1–5 described. Clinical improvement during somatostatin analog therapy is mainly mediated via a direct inhibitory effect on hormone production from the tumors, seen in 30%–70% of the patients. Also indirect non-tumor mediated effects on peripheral target organs contribute to the subjective improvement, achieved in 30%–70% of patients. Recently, significant improvement of quality of life has been demonstrated with long-acting depot formulations. There is little or no effect on tumor growth during octreotide therapy; tumor shrinkage has been reported in 10%–20% of patients, but stabilization of tumor growth can be achieved in about half of the patients with a duration of 8–16 months. Recently, induction of apoptosis has been described with high doses of lanreotide (12 mg/d). Eventually, however, all patients escape from somatostatin analog therapy with regard both to hormonal production and tumor growth, and the mechanism behind the tachyphylaxis is not yet known. Studies of optimal dosage and modes of administration, development of new slow release formulations, the potential value of high-dose somatostatin analog therapy and novel somatostatin receptor subtype specific analogs are important directions for the use of somatostatin analogs in the future. In addition, assessment of somatostatin receptor status for each patient and studies of tumor biology, e.g., inhibition of exocytosis, antiproliferative effects and induction of apoptosis during treatment will help to optimize treatment and provide new insights into mechanisms of action of somatostatin analogs.
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  • 16
    ISSN: 1433-044X
    Keywords: Schlüsselwörter ; HWS • Diskoligamentäre Instabilität • Funktionsaufnahmen • MRT ; Key words ; Lower cervical spine • Traumatic discoligamentous instabilities • Magnetic resonance imaging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary The diagnostic accuracy of magnetic resonance imaging (MRI) for traumatic discoligamentous instabilities of the lower cervical spine was evaluated. MRI findings were correlated with the intraoperative findings. Fifteen patients were included in the study (11 males, 4 females, 33 years old on average). Indication for surgery was established with dynamic a. p. instability under fluoroscopy. In the operation, all patients showed complete traumatic rupture of the intervertebral disc. In contrast, MRI sequences revealed only eight mild and four severe disc lesions. Intraoperatively ruptures of the anterior and posterior longitudinal ligaments were verified in 7 patients each. MRI studies depicted only five ruptures of the anterior and three ruptures of the posterior longitudinal ligaments. In three cases MRI demonstrated no pathological findings (3 of 15 false-negative results). MRI and intraoperative findings showed no statistical correlation. MRI is of only limited value in diagnosing traumatic discoligamentous instabilities of the lower cervical spine.
    Notes: Zusammenfassung Die diagnostische Aussagekraft der Magnetresonanztomographie (MRT) bei frischer, traumatischer diskoligamentärer Instabilität der unteren Halswirbelsäule (HWS) wurde untersucht. Hierzu wurden die Befunde der MRT-Diagnostik mit den intraoperativen Befunden korreliert. Die Indikation zur Operation wurde bei nachgewiesener translatorischer a.-p.-Instablität im Röntgenbildwandler bei dynamischer Untersuchung gestellt; 15 Patienten (11 Männer, 4 Frauen, Durchschnittsalter 33 Jahre) wurden in die Studie eingeschlossen. Bei allen Patienten lag intraoperativ eine Ruptur des Discus intervertebralis vor. In der MRT-Diagnostik wurden hingegen bei 8 Patienten eine Bandscheibenprotrusion und bei 4 Patienten ein Bandscheibenprolaps nachgewiesen. Das vordere Längsband und das hintere Längsband waren intraoperativ bei jeweils 7 Patienten rupturiert. Im MRT-Befund war das vordere Längsband bei 5 Patienten und das hintere Längsband bei 3 Patienten rupturiert. Bei 3 von 15 Patienten (20 %) war die MRT unauffällig und damit falsch-negativ. Zwischen intraoperativem und MRT-Befund war keine statistisch signifikante Korrelation nachweisbar. Die MRT bietet daher für die Indikationsstellung zur Operation zum jetzigen Zeitpunkt keine ausreichende diagnostische Sicherheit bei der Evaluation traumatischer diskoligamentärer Instabilitäten der unteren HWS.
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  • 17
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    Der Unfallchirurg 102 (1999), S. 975-978 
    ISSN: 1433-044X
    Keywords: Schlüsselwörter ; Implantatversagen • UTN • Entfernung des distalen Nagelanteils ; Key words ; Breakage of UTN • Extraction device
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary By using UTN in the treatment of very distal tibia fractures delayed fracture-healing and pseudarthrosis with breakage of the nail are described. The removal of the distal solid part of the nail shows a special difficulty. We report about two cases of broken UTN in 1997. A simple extraction device to remove the distal part of the broken nail is demonstrated.
    Notes: Zusammenfassung Mit der Anwendung des unaufgebohrten Tibianagels (UTN) bei sehr distalen Tibiafrakturen wird zunehmend von verzögerter Knochenbruchheilung und Pseudarthrosen mit nachfolgendem Implantatversagen berichtet. Die Entfernung des soliden, distalen Nagelteils bringt besondere Schwierigkeiten mit sich. 1997 wurden in der BG-Unfallklinik Ludwigshafen 2 gebrochene, unaufgebohrte Tibianägel entfernt. Der Behandlungsverlauf sowie ein einfaches, überall verfügbares Verfahren zur zuverlässigen Entfernung des distalen Nagelanteils werden beschrieben.
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  • 18
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    Monatsschrift Kinderheilkunde 147 (1999), S. 1014-1017 
    ISSN: 1433-0474
    Keywords: Schlüsselwörter Bloom-Syndrom ; Chromosomeninstabilität ; Kleinwuchs ; Wachstumshormon ; Key words ; Bloom’s syndrome ; Chromosomal instability ; Growth retardation ; Growth hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Bloom’s syndrome is a rare condition which is doubtlessly underdiagnosed. It is characterized by pre- and postnatal growth deficiency, cutaneous lesions, a recognizable facial dysmorphism, recurrent infections and an excessive risk of cancer. The pathogenetic basis is a significantly increased genomic instability. Bloom’s syndrome is transmitted as an autosomal-recessive trait. The responsible gene has been identified recently. We report on a 4 year old Turkish girl with Bloom’s syndrome confirmed by an increased sister chromatid exchange rate. She came to our attention because of severe growth deficiency, but lacked the cutaneous signs. Discussion: Our report emphasizes that Bloom’s syndrome has to be considered in children presenting with proportionate short stature, even in the absence of the full clinical phenotype.
    Notes: Zusammenfassung Das Bloom-Syndrom ist eine seltene, zweifellos aber auch zu selten diagnostizierte Entität. Es ist gekennzeichnet durch prä- und postnatale Wachstumsverzögerung, Hautveränderungen, eine diskrete, aber typische faziale Dysmorphie, gehäufte Infektionen und – prognostisch am bedeutsamsten – ein hohes Malignomrisiko. Der zugrundeliegende Defekt ist eine erhöhte DNA-Instabilität. Der Erbgang ist autosomal-rezessiv; das Gen ist seit kurzem bekannt. Wir berichten über ein 4jähriges türkisches Mädchen mit den Leitsymptomen Kleinwuchs und Dystrophie, bei dem die Diagnose anhand der erhöhten Schwesterchromatidaustauschrate gesichert werden konnte. Das Kind hatte nicht die typischen Hautveränderungen. Diskussion: Der Fallbericht verdeutlicht, daß das Bloom-Syndrom in die Differentialdiagnose des proportionierten Kleinwuchses einbezogen werden muß, auch wenn nicht das Vollbild der Erkrankung vorliegt.
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  • 19
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    Der Unfallchirurg 102 (1999), S. 918-923 
    ISSN: 1433-044X
    Keywords: Schlüsselwörter ; Fehlheilung • Posttraumatische Arthrose • Lisfranc-Luxationsfrakturen • Lisfranc-¶Arthrodese • Maryland-Foot-Score ; Key words ; Residual deformity • Posttraumatic arthritis • Lisfranc-fracture-dislocation • Lisfranc Arthrodesis • Maryland Foot Score
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary The causes of residual deformity with posttraumatic painful arthritis after tarsometatarsal (Lisfranc) fracture-dislocation with the need for correctional arthrodesis are in our experience (22 cases over 5 years) overseen injuries in one third, closed reduction and immobilisation or inadequate fixation technique with K-wires in another two thirds of cases. Foot malalignment and residual instability is assessed with weight-bearing radiographs of both feet, adduction/abduction stress films and CT scans for complex deformity. Correction is carried out via two longitudinal dorsal incisions, strict epiperiosteal preparation and debridement of the Lisfranc joint of all remaining cartilage, sclerosis and fibrous tissue. Reorientation begins, in the same manner as primary open reduction, with anatomical alignment of the second metatarsal base to the second cuneiform. Defects are filled with autologous bone grafting, stable fusion can be achieved with 3.5 mm cancellous compression screws. Full weight bearing is allowed in a modelled plaster shoe for 6 to 8 weeks. The functional medium-term results are convincing with 15 of 17 patients seen after 13 months of follow-up working full time. The Maryland Foot Score improved from 38.9 to 76.8 points in these patients.
    Notes: Zusammenfassung Fehlverheilungen mit konsekutiver posttraumatischer Arthrose nach Lisfranc-Luxationsfrakturen mit notwendiger reorientierender Lisfranc-Arthrodese gehen nach eigener Erfahrung (22 Fälle in 5 Jahren) in 1/3 der Fälle auf übersehene Verletzungen, in 2/3 der Fälle auf unzureichende geschlossene Repositionen und perkutane Spickdrahtosteosynthesen oder alleinige Retention im Gipsverband zurück. Belastungsaufnahmen beider Füße in 2 Ebenen sind zur Erkennung der Instabilität und Fehlverheilung unerläßlich, gegebenenfalls Vorfußabduktions- bzw. Adduktionsaufnahmen, eine Tomographie oder ein CT in 2 Ebenen. Bei veralteter homolateraler Lisfranc-Luxationsfraktur ist über 2 dorsale Längsinzisionen bei streng epiperiostaler Präparation das gesamte Lisfranc-Gelenk auszuräumen. Die Reorientierung beginnt – wie bei frischer Verletzung mit der anatomischen Einpassung der Metatarsale-II-Basis zum Cuneiforme II. Defekte sind mit autologer Spongiosa aufzufüllen, die Arthrodese mit 3,5er Kortikaliszugschrauben ist ausreichend. Die Nachbehandlung im Gipsschuh für 6–8 Wochen mit Vollbelastung hat sich bewährt. Der funktionelle Gewinn nach reorientierender Lisfranc-Arthrodese ist nach dem Maryland-Foot-Score von präoperativ 38,9 auf 76,8 Punkte postoperativ überzeugend.
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  • 20
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    Der Unfallchirurg 102 (1999), S. 972-974 
    ISSN: 1433-044X
    Keywords: Schlüsselwörter ; Juvenile Knochenzyste • Pathologische Fraktur • Konservative Therapie • Spontanheilung ; Key words ; Juvenile bone cyst • Pathological fracture • Conservative therapy • Spontaneous healing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Up to the present only few cases of spontaneous healing of juvenile bone cysts after pathological fractures were published. Most authors will not accept the possibility of a “healing fracture” in bone cysts because of missing documentation. In our clinic we could observe spontaneous healing of a juvenile bone cyst of the tibia after a pathological fracture in a ten-year old boy. Therefore healing of bone cysts after fracture is proven. On the other hand healing of bone cysts following conservative treatment after a fracture is not sure. Conservative treatment depends on the localization and type of the fracture. We recommend nonoperative treatment of first pathological fractures in small-sized bone cysts. This kind of treatment can be especially advised for young patients up to the tenth year because there are only few complications in fracture healing and a relatively high recurrence rate following operative treatment. In case of a refracture there are better conditions for fracture healing after surgery.
    Notes: Zusammenfassung Nach pathologischer Fraktur juveniler Knochenzysten (JKZ) wurde in der Literatur über einzelne Fälle von Spontanheilungen der Zysten im Rahmen der Frakturheilung berichtet. Vollständig dokumentierte Fälle liegen jedoch nicht vor, so daß die Möglichkeit einer sog. „heilenden Fraktur“ von den meisten Autoren nicht akzeptiert oder sogar prinzipiell ausgeschlossen wird. In unserer Klinik wurde eine Spontanheilung einer juvenilen Knochenzyste der Tibia nach pathologischer Unterschenkelfraktur bei einem 10 jährigen Jungen beobachtet. Eine Spontanheilung der JKZ im Rahmen der Frakturheilung ist also sicher grundsätzlich möglich. Eine zuverlässige Ausheilung der Zyste kann allerdings auf konservativem Wege nicht erreicht werden, da die osteolytische Potenz erhalten bleibt. Voraussetzung für ein konservatives Vorgehen ist eine geeignete Frakturlokalistation und -form. Unseres Erachtens ist die konservative Behandlung bei einer erstmaligen Spontanfraktur kleinerer Zysten indiziert. Insbesondere gilt dies für junge Patienten bis zum 10. Lebensjahr, aufgrund des günstigen Frakturheilungspotentials bei gleichzeitig bestehendem hohen Zystenrezidivrisiko trotz operativer Therapie. Im Falle der Refraktur ergeben sich für den dann notwendigen Eingriff evtl. günstigere Voraussetzungen aufgrund der Alterszunahme.
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  • 21
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    Der Unfallchirurg 102 (1999), S. 949-954 
    ISSN: 1433-044X
    Keywords: Schlüsselwörter ; Sprunggelenk • Begleitverletzungen •¶Fraktur • Prognose • Knorpelschaden ; Key words ; Ankle • Injury • Fracture • Outcome •¶Cartilaginous lesion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary The objective of this prospective study was to determine clinical and radiological results 2 years after ankle fractures and to compare them with preoperative arthroscopic findings. Of 135 consecutive patients undergoing osteosynthetic treatment, 128 (type A, 2; type B, 97; type C, 29) were reviewed 2 years later. The Kitaokascore for functional and clinical rating and the Kannusscore for radiological rating were used. Initial cartilaginous lesions localized on the medial malleolus led to poor clinical ratings (P 〈 0.01) and cartilaginous lesions of the pilon tibiale and talus caused poor radiological results (P 〈 0.032 and P 〈 0.046). After rupture of the deltoid ligament, more pain was observed (P 〈 0.038). Rupture of the syndesmosis, however, did not affect the outcome. Clinical outcome was poor among the 30- to 65-year-old patients. The clinical and radiological outcome of women was poorer (P 〈 0.018 and P 〈 0.046) than for men. The older the patients, the more radiological signs of arthritis were found. The outcome of ankle fracture seems to correlate significantly with the nature, dimension and localization of fracture-associated cartilaginous and ligament lesions. Age and sex also seem to play a significant role in the outcome.
    Notes: Zusammenfassung Die Ziele dieser prospektiven Studie war es, die klinischen und radiologischen Resultate 2 Jahre nach Malleolarfraktur zu bestimmen und diese mit den initialen, arthroskopisch erhobenen Begleitverletzungen nach dem Unfall zu vergleichen. Von 135 konsekutiven Patienten (135 Füsse), die vor der osteosynthetischen Versorgung einer Malleolarfraktur arthroskopisch untersucht wurden, konnten 128 Patienten (Typ A, 2; Typ B, 97; und Typ C, 29) nach durchschnittlich 2 Jahren nachkontrolliert werden. Dabei wurden der Kitaoka-Score (Klinik, Funktion) und der Kannus-Score (Röntgen) verwendet. Initiale Knorpelschäden am medialen Malleolus führten zu einem schlechteren klinischen Resultat (P 〈 0.010), wohingegen Knorpelschäden am Pilon tibiale und Talus ein schlechteres radiologisches Resultat (P 〈 0.032 bzw. P 〈 0.046) zeigten. Nach Ruptur des Lig. deltoideum waren assoziierte Schmerzen gehäuft (P 〈 0.038). Eine Syndesmosenruptur beeinflusste das Resultat nicht. Generell war das klinische Resultat bei den 30- bis 65 jährigen Patienten am schlechtesten und bei Frauen klinisch und radiologisch schlechter als bei Männern (P 〈 0.018 bzw. P 〈 0.046). Die radiologischen Veränderungen nahmen mit dem Alter zu. Das Resultat nach Malleolarfrakturen scheint demnach entscheidend von der Art, Ausmass und Lokalisation der Begleitverletzungen des Knorpels und der Bänder abzuhängen. Daneben könnten Alter und Geschlecht des Patienten präjudizierende Faktoren des Endergebnisses sein.
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  • 22
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    Der Unfallchirurg 102 (1999), S. 967-971 
    ISSN: 1433-044X
    Keywords: Schlüsselwörter ; Bakterielle Arthritis • Salmonella enteritidis • Hüftgelenk • Pathologische Diagnostik • Therapie ; Key words ; Bacterial arthritis • Salmonella enteritidis • Hip joint • Pathologic diagnosis • Therapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Secondary haematogenous joint involvement is seen in less than 1 % of patients with Salmonella infections. These atypical infections are frequently encouraged by pre-existing local or systemic disease. We present a case of a patient with known alcohol abuse who developed a septic infection of her right hip requiring resection of the femoral head. Histologic analysis showed signs of pre-existing osteonecrosis probably induced by alcohol intake. Cartilage and bone were invaded and destructed by an aggressive granulation tissue. Initially, a biopsy evaluated without knowledge of the positive bacteriological result had been interpreted as indicative of a tumour. The onset, clinical course, diagnosis and therapy of joint involvement by Salmonella are discussed with regard to our case and the relevant literature. This case illustrates the necessity for clinicians to share all their information about the patient with the pathologist.
    Notes: Zusammenfassung Sekundäre hämatogene Gelenkinfektionen werden bei weniger als 1 % aller systemischen Salmonellosen beobachtet. Lokale Gewebeschädigungen oder systemische Grunderkrankungen begünstigen das Zustandekommen dieser atypischen Infektionen. Wir stellen den Fall einer alkoholabhängigen Patientin dar, bei welcher eine Salmonellenkoxitis auf dem Boden einer ischämischen Knochennekrose des Hüftkopfes entstand. Die weit fortgeschrittene Zerstörung des Gelenks erforderte eine Resektionsarthroplastik. Histologisch fand sich eine destruierende Invasion von Knorpel und spongiösem Knochen durch zellreiches Entzündungsgewebe. In Unkenntnis des bakteriologischen Befundes war zunächst der Verdacht auf eine neoplastische Veränderung geäußert worden. Entstehung, Verlauf, Diagnostik und Therapie der Salmonellenarthritis werden anhand dieses Falles und der Literatur diskutiert. Die erfolgreiche Befundung einer Knochenbiopsie ist auf eine gute Kommunikation zwischen Kliniker und Pathologe angewiesen.
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  • 23
    ISSN: 1433-044X
    Keywords: Schlüsselwörter ; Arbeitsgemeinschaft „Wirbelsäule“ der DGU • Thorakolumbale Wirbelsäule • Wirbelsäulenverletzung • Operative Behandlung • Epidemiologie • Magerl-Klassifikation • Querschnittlähmung • Frankel-/ASIA-Score •¶Hannover Wirbelsäulenscore ; Key words ; Working group “spine“ of the German Society of Trauma Surgery • Thoracolumbar spine • Spinal fracture • Injuries of the spine • Operative treatment • Epidemiology • Classification of thoracolumbar spinal injuries by Magerl • Paraplegia • Frankel/ASIA grading system • Hannover Spine Score
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary The authors report on a prospective multicenter study with regard to the operative treatment of fractures and dislocations of the thoracolumbar spine. 18 traumatologic centers in Germany and Austria, forming the working group “spine“ of the German Society of Trauma Surgery, are participating in this continuing study. Between September 1994 and December 1996 682 patients (64 % male) with an average age of 391/2 (7–83) years were entered. The entry criteria included all patients with acute and operatively treated (within 3 weeks after trauma) fractures and dislocations of the thoracolumbar spine (Th 10-L 2). Part 1 of this publication outlines the protocol and epidemiologic data. The incidence of fractures and dislocations of the thoracolumbar spine and associated injuries were recorded according to a standardized protocol, as well as the different operative methods and complications, duration of hospital stay, rehabilitation and incapacity. The analysis of the clinical social and radiological course was a second focus. The most frequent mechanism of injury was a fall (50 %) or traffic accident (22 %). Most of the fractures occured at the L 1 level (49 %). All injuries were classified according to the ASIF (AO) classification. 65 % sustained an A-type fracture (compression fracture). Associated injuries were observed in 35 % and 6 % were polytraumatized. Extremities and thorax were most frequently affected. Younger age and traffic accidents lead more often to C-type fracture (fracture dislocation) and polytrauma. An increased number of multisegmental or multilevel lesions were observed in polytraumatized patients. There were 16 % with incomplete paraplegia (Frankel/ASIA B–D) and 5 % with complete paraplegia (Frankel/ASIA A). The rate of patients with initial neurologic deficits significantly increased with the severity of spinal injury according to the Magerl classification. Until discharge a neurologic improvement (at least 1 Frankel/ASIA grade) was observed in 32 % of the partially paralyzed (Frankel/ASIA B–D) and in 12 % of the patients with complete paraplegia (Frankel/ASIA A). A neurologic deterioration occured in 3 patients (0.4 %). As a base for further follow-up and late results the individual starting point was determined by collecting relevant data of the patients' history: 277 (40.6 %) patients suffered from simultaneous diseases, one half was spine related. At the time of injury 559 (82.0 %) patients were employed; 429 (62.9 %) doing manual work. 369 (54.1 %) patients stated sportive activities before the injury and 561 (82.3 %) designated their “back function“ as normal. For the time before injury the patients scored an average of 93.4 points in the Hannover Spine Score (0–100 points concerning complaints and function of the back/spine).
    Notes: Zusammenfassung Die Autoren berichten über eine prospektive, multizentrische Studie zur operativen Behandlung frischer Verletzungen des thorakolumbalen Übergangs der Wirbelsäule. An der noch nicht abgeschlossenen Studie sind insgesamt 18 unfallchirurgische Kliniken in Deutschland und Österreich beteiligt; sie bilden die Arbeitsgemeinschaft „Wirbelsäule“ der Deutschen Gesellschaft für Unfallchirurgie. Von September 1994 bis Dezember 1996 wurden 682 Patienten (64 % männlich) mit einem Durchschnittsalter von 391/2 (7–83) Jahren erfaßt. Die Eingangkriterien schlossen alle Patienten mit frischer und innerhalb von 3 Wochen operierter thorakolumbaler Verletzung (Th 10-L 2) ein. Im Teil 1 des Berichts schildern wir den Aufbau der Studie und epidemiologische Daten. Häufigkeit und Art von Wirbelsäulen- und Begleitverletzungen wurden ebenso einheitlich erfaßt wie Angaben zu den verschiedenen operativen Behandlungskonzepten und Komplikationen, der Dauer der Behandlung, Rehabilitation und Arbeitsunfähigkeit. Die Analyse des klinischen, sozialen und röntgenologischen Verlaufs stellte einen weiteren Schwerpunkt dar. Häufigste Unfallursachen waren Stürze aus der Höhe (50 %) und Verkehrsunfälle (22 %). Der erste Lendenwirbelkörper war mit Abstand am häufigsten betroffen (49 %). Alle Verletzungen wurden nach der AO-Einteilung klassifiziert; 65 % aller Patienten erlitten Kompressionsverletzungen (Typ A). Bei 59 % lag eine isolierte Wirbelsäulenverletzung vor, die übrigen wiesen Begleitverletzungen auf und 6 % waren polytraumatisiert. Begleitverletzungen betrafen am häufigsten Extremitäten und Thorax. Jüngere Patienten und Verkehrsunfallopfer erlitten häufiger eine Rotationsverletzung (Typ C) und waren häufiger polytraumatisiert. Mehrsegment- oder Mehretagenläsionen waren häufiger bei Polytraumatisierten. Eine inkomplette Querschnittläsion (Frankel/ASIA B–D) wiesen 16 %, eine komplette (Frankel/ASIA A) 5 % auf. Der Anteil neurologisch beeinträchtigter Patienten nahm mit der Verletzungsschwere signifikant zu. Bei 32 % der Patienten mit inkomplettter Lähmung besserte sich der neurologische Befund bis zur Entlassung um mindestens 1 Stufe nach dem Frankel-/ASIA-Schema, bei 12 % nach Paraplegie. Zu einer Verschlechterung um mindestens 1 Stufe kam es bei 3 Patienten (0,4 %). Als Grundlage differenzierter Nachuntersuchungsergebnisse wurden Angaben zur individuellen Situation vor dem Unfall erfaßt: 277 (40,6 %) Patienten wiesen Vorerkrankungen auf, etwa die Hälfte davon solche der Wirbelsäule. 559 (82,0 %) Patienten waren zum Zeitpunkt des Unfalls berufstätig; 429 (62,9 %) gingen einer körperlichen Arbeit nach. 369 (54,1 %) Patienten gaben sportliche Freizeitaktivitäten an und 561 (82,3 %) bezeichneten ihre Rückenbeweglichkeit als normal. Im Hannover Wirbelsäulen-Score (0–100 Punkte zu Beschwerden und Funktion) erzielten die Patienten für die Zeit vor dem Unfall im Mittel 93,4 Punkte.
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  • 24
    ISSN: 1436-2813
    Keywords: Key Words: nitric oxide ; DNA damage ; apoptosis ; tumor necrosis factor-α ; mitochondrial respiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: S -nitroso-N-acetylpenicillamine protected cul-tured L929 cells from apoptosis induced by tumor necrosis factor-α (TNF-α) plus actinomycin D, as determined by the detection of DNA fragmentation and morphological changes. NO also prevented an enhancement of the production of reactive oxygen intermediates by TNF-α plus actinomycin D, as assessed by the oxidation of dihydrorhodamine 123 and hydroethidine. Because the inhibition of mitochondrial respiration by rotenone or antimycin A suppressed the increased oxidation of both dihydrorhodamine 123 and hydroethidine, it was suggested that TNF-α accelerated the leakage of reactive oxygen intermediates from the mitochondrial electron transport system. Polarography showed that NO reversibly inhibited mitochondrial respiration at either complexes I–III, II–III, or IV, thus suggesting the inhibition of cytochrome oxidase. Taken together, these findings indicate that the decreased mitochondrial formation of reactive oxygen intermediates in the presence of NO might have a protective effect against TNF-α plus actinomycin D-induced apoptosis.
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  • 25
    ISSN: 1436-2813
    Keywords: angiogenesis ; p53 ; apoptosis ; lung adenocarcinoma ; hematogenous metastasis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The purpose of this study was to clarify which factors are important as predictors not only of patient survival but also of hematogenic metastasis in 15 patients with stage I lung adenocarcinoma who underwent curative operation. The relationship between tumor angiogenesis, apoptosis, and p53 oncogene was also studied. A total of 15 patients were divided into two groups: surviving group (n=7) and nonsurviving (metastasis) group (n=8). We studied the medical charts, operative records, pathologic reports, and tumor specimens taken at surgical resection. We measured the apoptotic index using the ApopTag kit and the intratumoral microvessel count using an anti-CD34 monoclonal antibody. In addition, immunohistochemical staining for the expression of p53 was conducted simultaneously. The clinicopathological characteristics, including age, sex, tumor size (pT), and histological differentiation, were not significantly different between the surviving and the nonsurviving group. The microvessel count was significantly higher in nonsurviving group than in the surviving group. The apoptotic index and the expression of p53 was not significantly different between the two groups. An inverse correlation between the apoptotic index and microvessel count, and a positive correlation between the expression of p53 and microvessel count, were observed. Angiogenesis may be an important prognostic factor in patients with stage I lung adenocarcinoma.
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  • 26
    ISSN: 1436-2813
    Keywords: Key Words: angiogenesis ; p53 ; apoptosis ; lung adenocarcinoma ; hematogenous metastasis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: n = 7) and nonsurviving (metastasis) group (n = 8). We studied the medical charts, operative records, pathologic reports, and tumor specimens taken at surgical resection. We measured the apoptotic index using the ApopTag kit and the intratumoral microvessel count using an anti-CD34 monoclonal antibody. In addition, immunohistochemical staining for the expression of p53 was conducted simultaneously. The clinicopathological characteristics, including age, sex, tumor size (pT), and histological differentiation, were not significantly different between the surviving and the nonsurviving group. The microvessel count was significantly higher in nonsurviving group than in the surviving group. The apoptotic index and the expression of p53 was not significantly different between the two groups. An inverse correlation between the apoptotic index and microvessel count, and a positive correlation between the expression of p53 and microvessel count, were observed. Angiogenesis may be an important prognostic factor in patients with stage I lung adenocarcinoma.
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  • 27
    Electronic Resource
    Electronic Resource
    Springer
    Arthroskopie 12 (1999), S. 305-312 
    ISSN: 1434-3924
    Keywords: Schlüsselwörter ; Kreuzbandersatzoperation ; Hamstring-Sehnen ; Femorale Fixation ; Tibiale Fixation ; Key words ; Anterior cruciate ligament reconstruction ; Hamstrings ; Femoral fixation ; Tibial fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: In recent years ACL reconsruction with the midpatellar tendon has been being called into question more and more because of high morbidity at the harvest site and problems with fixation near the joint. The”wiper effect” or “bungee-effect” may be mentioned as an example. In this paper, ACL reconstruction using the hamstrings is presented, which, in contrast to fixation with the Endobutton, has a very high pull out strength and also tibial fixation near the joint with a biodegradable interference screw. This surgical method has been carefully developed and is technically sophisticated. The technique can be used mainly in younger women with small patellae, athletically inactive persons with a high level of instability, all patients with parapatellar problems or pain, and patients whose occupation involves kneeling. The short-term results are excellent; long-term results with this new method are not yet available. This method should be seen as an alternative to ACL reconstruction with the midpatellar tendon and, in our view, the security of fixation is at least as good as with the Endobutton.
    Notes: In den letzten Jahren wird die Kreuzbandersatzoperation mit dem mittleren Patellarsehnendrittel zunehmend mehr in Frage gestellt. Gründe sind die hohe Transplantatentnahmemorbidität sowie Probleme bei der gelenknahen Fixation. Der Scheibenwischereffekt, der Bungee-Effekt u. a. seien hierfür stellvertretend genannt. Im folgenden wird daher eine Kreuzbandersatzoperationstechnik unter Verwendung der Hamstring-Sehnen vorgestellt, die im Gegensatz zur Fixation über den Endobutton ® eine sehr ausreißfeste femorale Fixation sowie eine ebenfalls gelenknahe tibiale Fixation über eine biodegradierbare Interferenzschraube beinhaltet. Die Operationsmethode ist durchdacht und technisch ausgereift. Indikationen stellen vorwiegend jüngere Frauen mit kleiner Kniescheibe, sportlich wenig aktive Menschen mit hochgradiger Instabilität sowie alle Patienten mit parapatellaren Problemen oder Schmerzen, des weiteren Patienten mit kniender Tätigkeit dar. Die Kurzzeitresultate sind hervorragend, Langzeitergebnisse stehen bei dieser jungen Methode noch aus. Die Methode sollte als Alternativvariante zur Kreuzbandersatzoperation mit dem mittleren freien Patellarsehnendrittel gesehen werden und ist in ihrer Fixationssicherheit aus unserer Sicht dem Endobutton mindestens gleichwertig.
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  • 28
    ISSN: 1434-3924
    Keywords: Schlüsselwörter ; Vorderes Kreuzband ; Rekonstruktion ; Patella-bone-tendon-bone ; Press-fit-Fixation ; All-Press-fit ; Doppelblockfixierung ; Key words ; ACL Reconstruction ; Bone-Patellar-tendon-bone autograft ; Press-fit ; Implant free fixation ; All-Press-Fit-fixation ; Doubleblock fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: In the treatment of anterior cruciate ligament reconstruction, bone-patellar-tendon-bone allograft has been developed to a standard. Fixation of the graft with interference screws in the femur and/or with staple in the tibia are usual. ALL-PRESS-FIT fixation makes an implant-free acl-reconstruction possible. The bone-patellar-tendon-bone allograft is harvested with oversized bone blocks. The “Bone-Compactor” is used for modelling the oversized boneblocks for press-fit-fixation in femur und tibia. Implantation can be managed in either an arthroscopic or open procedure. The proximal boneblock is implanted with a sagittal alignment of the tendon. While rotating the distal block 90° to medial, a recontruction of anteromedial und posterolateral fascicle is possible. Tension can be controlled after step-by-step fixation proximal and distally with a distalisation of the tibial block at the end. The prominent tibial block will be pressed in on corticalis-niveau with the “Reconcillation-Hit”. A femoral or tibial “Doubleblock-Fixation” allows a better anatomical reconstruction. A third bone block, harvested from the tibial channel, fixed on the distal ligament, is used to fill up the tibial channel with bone to the tibial plateau. The “tibial doubleblock-fixation” allows a better biomechanical bone fixation of the ligament. Alternatively, a patellar-bone-tendon-bone allograft can be used with a high tibial press-fit fixation with a ligament outlet directly onto the tibia plateau. The proximal boneblock is implanted deep in the whole femur whole and fixed with the third bone block harvested from the tibial channel.
    Notes: Bei der operativen Versorgung der vorderen Kreuzbandruptur hat sich die Verwendung des mittleren Drittels der Patellarsehne als Standard etabliert. Die Knochenblöcke aus Patella und Tibiakopf werden üblicherweise mit Interferenzschrauben oder Staple fixiert. Mit der All-Press-FIT-Verankerung ist eine fremdimplantatfreie femorale und tibiale Verankerung möglich. Der mittlere Sehnenanteil des Lig. patellae wird mit Knochenblöcken aus der distalen Patella und dem Tibiakopf gewonnen. Mit dem Knochenkompaktor, der als Zurichtungsinstrument und Schablone dient, werden die Knochenblöcke in notwendiger Übergröße so zugerichtet, daß sie später femoral und tibial ohne zusätzliche Hilfsmittel press fit verankert werden können. Die Implantation ist sowohl arthroskopisch als auch offen möglich. Der obere Knochenblock wird so eingebracht, daß ein vorderes und hinteres Bündel entsteht. Durch Rotieren des unteren Knochenblocks entsteht ein anteromediales und posterolaterales Bündel. Zur nachträglichen Korrektur der Bandspannung kann der tibiale Knochenblock nach distal ausgetrieben werden. Der Knochenblock wird dann mit dem „Versöhnungsschlag“ auf Kortikalisniveau versenkt. Die femorale oder tibiale „Doppelblockfixierung“ ermöglicht eine anatomiegerechte Rekonstruktion. Ein aus dem Tibiakopf entnommener dritter kortikospongiöser Knochenblock armiert den Ligamentanteil im Tibiakanal für eine bessere knöcherne Führung des Bands. Die so erzielte tibiale Doppelblockfixierung ermöglicht eine anatomiegerechte sagittale Ausrichtung des Ligaments durch eine langstreckige knöcherne Stabilisierung im Tibiaplateau. Alternativ wird der tibiale Knochenblock auf Höhe des Tibiaplateau press fit festgesetzt. Der patellare Knochenblock wird tief in dem femoralen Kanal eingesetzt. Das Ligament wird mit dem Eintreiben des aus dem Tibiabohrkanal gewonnenen Knochenblocks angespannt und femoral gelenknah verankert. Es entsteht die femorale Doppelblockfixierung.
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  • 29
    ISSN: 0340-1855
    Keywords: Key words ; Rheumatoid arthritis ; DMARD therapy ; observation study ; Schlüsselwörter ; Rheumatoide Arthritis ; Basistherapie ; Langzeitverlauf
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Ziel der Arbeit: Untersuchung der Langzeiteffektivität und Verträglichkeit der Basistherapie mit Methotrexat (MTX), Azathioprin (AZA), intramuskulärem Gold, D-Penicillamin (DPA), Sulfasalazin (SASP) und Cholorochin (CQ) bei rheumatoider Arthritis (RA). Methoden: Zwischen 1979 und 1994 wurden Daten von 1681 Patienten des Rheumazentrums Rostock prospektiv erfaßt. Eine 50%ige Reduktion der Anzahl geschwollener Gelenke war erforderlich, um die Therapie fortzuführen. Zusätzlich wurden in halbjährlichen Abständen ein modifizierter Lansbury-Index, der Bewegungsfunktionstest nach KEITEL sowie akute-Phase-Parameter erfaßt. Nebenwirkungen, die zum Abbruch der Therapie führten, wurden ebenfalls festgehalten. Ergebnisse: Nach einer Beobachtungszeit von mindestents vier Jahren nahmen noch 39,6% bzw. 28,3% der Patienten MTX bzw. AZA ein. 18,2% bekamen Gold, 16,9% DPA. SASP und CQ wurden von 13,5 bzw. 6,6% der Patienten eingenommen. Die Abbruchraten wegen Nebenwirkungen (NW) betrugen für MTX, AZA and SASP 15,9%, 15,3% bzw. 17,7%, während 34,8% CQ wegen NW absetzten (Gold: 27,4%, DPA: 26,9%). Die Mehrzahl der Therapieabbrüche erfolgte bereits im ersten Jahr der Behandlung. Subgruppen seropositiver Patienten bzw. Patienten mit Rheumaknoten oder erosiven Verläufen hatten ein schlechteres Ansprechen auf die Behandlung, unabhängig von der Art des eingesetzten Basistherapeutikums. Schlußfolgerung: MTX war in der Langzeittherapie die effizienteste Substanz, gefolgt von AZA, während CQ die geringste Rate von Langzeitanwendern aufwies. Die Ergebnisse unterstreichen den Stellenwert von Langzeitbeobachtungen unter klinischen Alltagsbedingungen in Ergänzung zu kontrollierten klinischen Studien.
    Notes: Summary Objective: To study the long-term efficacy and safety of methotrexate (MTX), intramuscular gold, azathioprine (AZA), chloroquine (CQ), sulphasalazine (SASP), and D-penicillamine (DPA) in rheumatoid arthritis (RA) patients. Methods: Between 1979 and 1994, clinical data were prospectively gathered in a single center. 1681 patients were followed-up for at least 4 years. A 50% reduction of the swollen joint count was required to continue therapy. In addition, a modified Lansbury index, the Keitel function test, and laboratory parameters were determined every six months. Side effects leading to the discontinuation of treatment were recorded as well. Results: After an observation period of more than four years, 39.6% and 28.3% of patients were taking MTX and AZA, respectively; 18.2% were receiving gold, 16.9% remained on DPA. SASP and CQ were still applied in 13.5% and 6.6%. MTX, AZA and SASP had a drop-out rate due to toxicity of 15.9%, 15.3% and 17.7%, whereas 34.8% had to discontinue CQ (gold: 27.4%, DPA: 26.9%). The majority of dropouts occurred within the first year of treatment. Subgroups of seropositive patients and patients with rheumatoid nodules had a poorer treatment efficacy irrespective of the DMARD. Conclusion: In the long-term application, MTX was the most efficient compound, followed by AZA, whereas CQ had the poorest drug survival. Our results underline the value of long-term observations under the conditions of clinical practice as a supplement to controlled clinical trials.
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  • 30
    Electronic Resource
    Electronic Resource
    Springer
    Diabetologia 42 (1999), S. 711-718 
    ISSN: 1432-0428
    Keywords: Keywords Malnutrition ; ageing ; beta-cell mass ; apoptosis ; glucose tolerance.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Aims/hypothesis. In a recently developed rat model, maternal food restriction from day 15 of pregnancy until weaning induced low birth weight and a 70 % reduction of beta-cell mass in the offspring at day 21 after birth. Subsequent renutrition from weaning was insufficient to fully restore beta-cell mass in young adult rats. The aim of this study is to investigate the long-term consequences of early malnutrition on beta-cell mass and function. Methods. Oral glucose tolerance tests were done in 3- and 12-month-old animals and beta-cell mass and apoptosis were determined by morphometrical measurements on pancreatic sections. The specific impact of postnatal malnutrition was studied by comparing control animals (C group) with animals malnourished during their fetal life only (R/C group), and animals malnourished during fetal life and until weaning (R group). Results. In 3-month-old R/C animals beta-cell mass reached 8.0 ± 1.5 mg with no further increase until 12 months (8.1 ± 1.5 mg), compared with 9.3 ± 1.9 mg in control rats. Twelve-month-old R/C animals showed normal plasma insulin responses and borderline glucose tolerance. In R animals, apoptosis reached 1.9 ± 0.4 % of the beta cells at 3 months, compared with 0.7 ± 0.5 % in control rats, and beta-cell mass did not increase between 3 and 12 months (4.7 ± 0.8 mg at 12 months). In aged control and R animals, apoptosis affected 8 % of the beta cells. At 12 months only, R animals showed profound insulinopenia and marked glucose intolerance. Conclusion/interpretation. In conclusion, perinatal malnutrition profoundly impairs the programming of beta-cell development. In animals with decreased beta-cell mass the additional demand placed by ageing on the beta cells entails glucose intolerance since beta-cell mass does not expand and apoptosis is increased. [Diabetologia (1999) 42: 711–718]
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  • 31
    ISSN: 1432-0428
    Keywords: Keywords Type I diabetes ; interferon-γ ; transgenic mice ; apoptosis ; insulin secretion ; tumour necrosis factor.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Aims/hypothesis. To examine whether interferon-γ destroys islet beta cells directly or indirectly through lymphocyte activation, or whether direct action of interferon-γ on beta cells by itself induces diabetes without insulitis. Methods. To avoid possible nonspecific breakdown of beta cells by transgenic overexpression of interferon-γ by the insulin promoter, we generated transgenic mice expressing interferon-γ under the control of rat glucagon promoter (RGP-IFN-γ-Tg mice). Results. The absence of insulitis in RGP-IFN-γ-Tg mice enabled us to investigate the direct effects of paracrine interferon-γ. In RGP-IFN-γ-Tg mice, serum concentrations of interferon-γ and tumour necrosis factor-α (TNF-α) were 50 and 6 times higher than those in their littermates, respectively, and glucose-responsive insulin secretion decreased to one-half the level of that in the littermates. Transgenic interferon-γ induced remodelling of beta cells where apoptosis of many beta cells was compensated by their vigorous regeneration and diabetes did not occur in most of the RGP-IFN-γ-Tg mice. Conclusion/interpretation. Interferon-γ alone is insufficient for the complete destruction of beta cells in vivo, and factors other than interferon-γ including activated lymphocytes or other cytokines, are necessary in addition to interferon-γ for the development of Type I (insulin-dependent) diabetes mellitus. [Diabetologia (1999) 42: 566–573]
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  • 32
    ISSN: 1432-0428
    Keywords: Keywords Nicotinamide ; cytokine ; islet ; insulin ; apoptosis ; diabetes.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Nicotinamide intervention trials are presently undertaken to prevent Type I (insulin-dependent) diabetes in high risk subjects. They are based on studies in rodents reporting nicotinamide protection against beta-cell injury in vitro and in vivo. This study examines whether nicotinamide can protect human beta cells in vitro. At concentrations (2 and 5 mmol/l) to protect rat beta cells against necrosis by streptozotocin or hydrogen peroxide, nicotinamide prevents hydrogen peroxide-induced necrosis of human beta cells. As with rat beta cells, nicotinamide fails to protect human beta cells against apoptosis induced by a combination of the cytokines interleukin-1β , interferon-γ and tumour necrosis factor-α. In rat beta cells, nicotinamide (2 to 20 mmol/l) was also found to induce apoptosis, in particular during the days following its protection against necrosis; this cytotoxic effect was not observed with human beta cells. These data demonstrate that nicotinamide can protect human beta cells against radical-induced necrosis, but not against cytokine-induced apoptosis. This effect is not associated with a delayed apoptosis as in rat beta cells. [Diabetologia (1999) 42: 55–59]
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  • 33
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    Der Radiologe 39 (1999), S. 52-59 
    ISSN: 1432-2102
    Keywords: Schlüsselwörter ; Key words
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary The etiology of chronic diseases of the ankle joint comprises a wide spectrum including chronic inflammatory processes and chronic degenerative, tumorous and neuropathic processes, as well as some specific syndromes based on chronic changes of the ankle joint. Of the inflammatory processes, chronic juvenile arthritis (JVC) is the most common disease. However, also Reiter disease, psoriasis or chronic monoarthritid diseases such as gout, as well as granulomatous diseases (tuberculosis, sarcoidosis) and fungal infections, may affect the ankle joint in a chronic course. Chronic degenerative changes are usually secondary due to abnormal positioning of the joint constituents or repetitive trauma. Neuropathic changes, as frequently seen in the course of diabetes, present with massive osseous destruction and malposition of the articular constituents. Chronic osseous as well as cartilaginous and synovial changes are seen in hemophilic patients. Chronic traumatic changes are represented by pigmented villonodular synovitis (PVNS), and chondromatosis, both with a predilection for the ankle joint. Due to the possibilities of magnetic resonance imaging (MRI), diagnosis of chronic ankle changes includes chronic ligamentous, tendinous and soft tissue changes. With the use of MRI, specific syndromes can be defined which particularly affect the ankle joint in a chronic way, such as the os trigonum syndrome, the anterolateral impingement syndrome and the sinus tarsi syndrome. Nevertheless, plain film radiographs are still the basic element of any investigation. MRI, however, can be potentially used as a second investigation, saving an unnecessary cascade of investigations with ultrasound and CT. The latter investigations are used only with very specific indications, for instance CT for subtle bone structures and sonography for a limited investigation of tendons or evaluation of fluid. Particularly due to the possibilities of MRI and the development of special gradient-echo imaging or high-resolution coils, the investigation of the ankle joint still offers a wide spectrum of innovation for the next years, which is particularly enforced by the increasing demand for specific diagnosis of chronic diseases in orthopedic medicine.
    Notes: Zusammenfassung Die Ätiologie chronischer Krankheitsbilder am Sprunggelenk ist vielfältig. Chronisch entzündliche Veränderungen, wie sie im Rahmen einer chronisch juvenilen Arthritis, des M. Reiter, der Psoriasis oder chronischer Monoarthritiden, wie der chronischen Gicht zu beobachten sind, haben nicht selten ihre Manifestation am Sprunggelenk. Seltenere entzündlich chronische Affektionen hingegen können jedoch auch bei enteropathischen Arthritiden, granulomatösen Erkrankungen, wie der Tuberkulose oder Sarkoidose, beobachtet werden. Auch chronisch verlaufende Pilzaffektionen können zu atypischen Bildern führen. Chronisch degenerative Erkrankungen entstehen am Sprunggelenk fast ausschließlich sekundär. Prädisponierend sind hier angeborene oder erworbene Fehlhaltungen sowie chronische Veränderungen auf Basis abgelaufener oder repetitiver Traumen. Der neuropathische Formenkreis stellt eine weitere Causa chronischer Veränderungen dar, mit im fortgeschrittenem Stadium massiven Gelenks- und ossären Veränderungen. Chronische Veränderungen sind auch bei der Hämophilie im Rahmen des Blutergelenkes zu erwarten. Als chronisch tumoröse Veränderungen können die pigmentierte villonoduläre Synovitis (PVNS) und die Gelenkschondromatose betrachtet werden, mit teils ossären und teils charakteristischen Kapsel und Weichteilveränderungen. Die modernen Gelenkdiagnostik umfaßt zudem auch chronische Veränderungen an den Sehnen und Bändern. Besondere diagnostische Herausforderungen stellen letzendlich Syndrome dar, die klinisch zwar beobachtet, jedoch erst durch moderne Bildgebung in den letzten Jahren zunehmend spezifiziert werden konnten. Diese umfassen zum Beispiel das „Os-trigonum-Syndrom”, das „anterolaterale Weichteilimpingement” und das „Sinus-tarsi-Syndrom”. Wie in der allgemeinen Gelenkdiagnostik ist auch weiterhin das Nativbild als Basisuntersuchung unerläßlich. Der Einsatz der Magnetresonanztomographie (MRT) als Zweituntersuchung erspart oft eine kostenintensive und belastende Untersuchungskaskade. Der Einsatz moderner Sequenzen oder gezielte Kontrastmittelgabe erlauben zunehmend spezifische Diagnosen. Der Einsatz der Sonographie und ebenso der CT muß gezielt durchgeführt werden. Die Vielfalt der möglichen chronisch pathologischen Veränderungen und dem gegenüberstehend die Vielfalt der Untersuchungsmethoden, im speziellen der MRT-Techniken, zusammen mit den komplexen anatomischen Verhältnissen am Sprunggelenk stellen eine besondere Herausforderung an den Radiologen dar.
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  • 34
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    Annals of oncology 10 (1999), S. 139-150 
    ISSN: 1569-8041
    Keywords: apoptosis ; chemotherapy ; clinical trials ; gene therapy ; p53
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Nearly twenty years after the initial discovery of p53, we are now in an ideal position to exploit our vast knowledge of p53 biology in the creation of novel cancer therapies. Disruption of p53 function through mutation, or other means, occurs very frequently in human cancer. Loss of p53 function has been linked with unfavourable prognosis in a large number of tumour types, as indicated by more aggressive tumours, early metastasis and decreased survival rates. Many different avenues of research have converged upon p53 to highlight this protein as being one of the foremost cellular responders to stress, in particular to DNA damage. Huge advances have been made in understanding the complex role p53 plays in the regulation of apoptosis and cell cycle arrest. This review is not meant to be a comprehensive description of p53 biology, but rather serves to highlight current progress in the development of p53- oriented cancer therapies. These may be categorised into three basic strategies: gene replacement therapy using wild-type p53, restoration of p53 function by other means and, finally, targeting of the p53 dysfunction itself. Rapid progress is expected to be made regarding the identification of conventional pharmaceutical agents which either work in a p53-independent manner or act preferentially in p53 defective cells. Gene replacement therapy with wild-type p53 also holds considerable potential for obtaining clinically relevant results quickly. The other forms of cancer therapies based around p53 are much further behind in the developmental process, but may prove to more efficacious in the long run, especially in terms of specificity. As with many other fields, the innovation of successful p53-oriented cancer therapies is only limited by our understanding of p53 biology and the creative use of such knowledge.
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  • 35
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    Annals of oncology 10 (1999), S. 122-126 
    ISSN: 1569-8041
    Keywords: apoptosis ; biliary carcinogenesis ; cholangiocarcinoma ; genotoxicity ; risk factors ; therapeutic strategies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Carcinomas of the biliary tract are rare cancers developing from the epithelial or blast-like cells lining the bile ducts. A variety of known predisposing factors and recent experimental models of biliary carcinogenesis (e.g., infection with the liver fluke Opisthorchis viverrini, models of chemically induced carcinogenesis and experimental models of pancreaticobiliary maljunction) have elucidated different stages of this complex system of biliary tumorigenesis. Chronic inflammatory processes, generation of active oxygen radicals, altered cellular detoxification mechanisms, activation of oncogenes, functional loss of tumor-suppressor genes and dysregulation of cell proliferation and cell apoptotic mechanisms have been identified as important contributors in the development of cholangiocarcinomas. In this review, the known mechanisms involved in the carcinogenesis of biliary epithelium are addressed. We will divide the topic into four stages: 1) Predisposition and risk factors of biliary cancer, 2) Genotoxic events and alterations leading to specific DNA damage and mutation patterns. 3) Dysregulation of DNA repair mechanisms and apoptosis, permitting survival of mutated cells and 4) Morphological evolution from premalignant biliary lesions to cholangiocarcinoma. Finally, established and hypothetical future therapeutic strategies directed towards specific pathogenetic events during biliary carcinogenesis will be addressed.
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  • 36
    ISSN: 1569-8041
    Keywords: acute myelogenous leukemia ; apoptosis ; ara-CTP ; cytosine arabinoside ; HL-60 ; retinoic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Accumulation of the cytosine arabinoside (ara-C) metabolite ara-C-triphosphate (ara-CTP) in leukemic blast cells is considered to be the main determinant of ara-C cytotoxicity in vitro and in vivo. Retinoids such as all-trans-retinoic acid (ATRA) have been shown to increase the sensitivity of acute myelogenous leukemic (AML) blast cells to ara-C. To investigate the mechanism of this sensitisation, the hypothesis was tested that ATRA augments cellular ara-CTP levels in human-derived myelogenous leukemia HL-60 cells. Materials and methods: The effect of ATRA and 13-cis-retinoic acid on ara-CTP accumulation and ara-C-induced apoptosis was studied. Ara-CTP levels were measured by high-performance liquid chromatography (HPLC), cytotoxicity by the tetrazolium (MTT) assay, and apoptosis by occurrence of DNA fragmentation (gel electrophoresis), cell shrinkage and DNA loss (flow cytometry). Results: Pretreatment of HL-60 cells with ATRA (0.01–1 µM) caused a significant decrease in intracellular ara-CTP levels; e.g., incubation for 72 hours with ATRA 1 µM prior to one hour ara-C 10 µM reduced ara-CTP levels to 41% ± 4% of control. Similar results were obtained after preincubation with 13-cis-retinoic acid. In spite of decreased ara-CTP levels, the cytotoxicity of the combination was supraadditive and ATRA augmented ara-C-induced apoptosis. Conclusions: At therapeutically relevant concentrations ATRA increased ara-C cytotoxicity and ara-C induced apoptosis but this augmentation is not the corollary of elevated ara-CTP levels. The feasibility of ara-C treatment optimisation via strategies other than those involving elevation of ara-CTP levels should be investigated further.
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  • 37
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    Molecular and cellular biochemistry 196 (1999), S. 13-21 
    ISSN: 1573-4919
    Keywords: apoptosis ; DNA fragmentation ; GSHPx-1 knockout mice ; GSHPx-1 transgenic mice ; ischemia/repurfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Apoptosis, a genetically controlled programmed cell death, has been found to play a role in ischemic reperfusion injury in several animal species including rats and rabbits. To examine whether this is also true for other animals, an isolated perfused mouse heart was subjected to 30 min of ischemia followed by 2 h of reperfusion. Experiments were terminated before ischemia (baseline), after ischemia, and at 30, 60, 90 and 120 min of reperfusion. At the end of each experiment, hearts were processed for the evaluation of apoptosis and DNA laddering. The in situ end labeling (ISEL) technique was used to detect apoptotic cardiomyocyte nuclei while DNA laddering was evaluated by subjecting the DNA obtained from the cardiomyocytes to 1.8% agarose gel electrophoresis followed by photographing under UV illumination. The results of our study revealed that apoptotic cells appear only after 60 min of reperfusion as demonstrated by the intense fluorescence of the immunostained genomic DNA when observed under fluorescence microscopy. None of the ischemic hearts showed any evidence of apoptosis. These results were corroborated with the findings of DNA fragmentation showing increased ladders of DNA bands in the same reperfused hearts representing integer multiples of the internucleosomal DNA length (about 180 bp). Since our previous studies showed a role of glutathione peroxidase (GSHPx) in apoptotic cell death, we performed identical experiments using isolated hearts from GSHPx-l knockout mice and transgenic mice overexpressing GSHPx-l. GSHPx-l knockout mice showed evidence of apoptotic cell death even after 30 min of reperfusion. Significant number of apoptotic cells were found in the cardiomyocytes as compared to non-transgenic control animals. To the contrary, very few apoptotic cells were found in the hearts of the transgenic mice overexpressing GSHPx-l. Hearts of GSHPx-l knockout mice were more susceptible to ischemia/reperfusion injury while transgenic mice overexpressing GSHPx- 1 were less susceptible to ischemia reperfusion injury compared to non-transgenic control animals. The results of this study clearly demonstrate a role of GSHPx in ischemia/reperfusion-induced apoptosis in mouse heart.
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  • 38
    ISSN: 1573-4919
    Keywords: MKP-1 ; Fas ligand ; Fas ; apoptosis ; prostate cancer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Recent studies have suggested that MAP kinase phosphatase 1 (MKP-1) is overexpressed in prostate cancer. To evaluate the role of MKP-1 in regulating cell death and tumor growth in prostate cancer, MKP-1 was conditionally overexpressed in the human prostate cancer cell line DU145. Overexpression of MKP-1 in DU145 cells blocked activation of stress-activated protein kinase (SAPK/JNK). MKP-1 overexpression in DU-145 cells was also found to inhibit Fas ligand (FasL)-induced apoptosis, as well as block the activation of caspases by Fas engagement. In addition, MKP-1 blocked the activation of apoptosis by transfected MEKK-1 and ASK-1, presumably through its inhibition of the SAPK/JNK family of enzymes. MKP-1 blocked the ability of FasL to induce loss of mitochondrial transmembrane potential (Δγm), suggesting that MKP-1 acts upstream of mitochondrial pro-apoptotic events induced by FasL and that the SAPK/JNK pathway may form the signaling link between Fas receptor and mitochondrial dysfunction. Thus, MKP-1 overexpression in prostate cancer may play a role in promoting prostate carcinogenesis by inhibiting FasL-induced cell death.
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  • 39
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    Molecular and cellular biochemistry 199 (1999), S. 125-137 
    ISSN: 1573-4919
    Keywords: apoptosis ; ADP-ribosylation ; caspases ; PARP ; PARG
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Poly(ADP-ribosylation) is a post-translational modification playing a relevant role in DNA damage recovery, DNA replication and viral integration. Several reports also suggest a modulation of this process during cell death by apoptosis. The aim of this review is to discuss the possible involvement of poly(ADP-ribosylation) during apoptosis, by dealing with general considerations on apoptosis, and further examining the correlation between NAD consumption and cell death, the regulation of poly(ADP-ribose) metabolism in apoptotic cells, the effect of poly(ADP-ribose) polymerase inhibition on cell death occurrence and the use of enzyme cleavage as a marker of apoptosis. Finally, the future prospects of the research in this area will be addressed.
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  • 40
    ISSN: 1573-4919
    Keywords: DNA binding protein ; NAD metabolism ; cellular response to DNA damage ; γ-rays ; alkylating agents ; genomic instability ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract A dual approach to the study of poly (ADP-ribose)polymerase (PARP) in terms of its structure and function has been developed in our laboratory. Random mutagenesis of the DNA binding domain and catalytic domain of the human PARP, has allowed us to identify residues that are crucial for its enzymatic activity. In parallel PARP knock-out mice were generated by inactivation of both alleles by gene targeting. We showed that: (i) they are exquisitely sensitive to γ-irradiation, (ii) they died rapidly from acute radiation toxicity to the small intestine, (iii) they displayed a high genomic instability to γ-irradiation and MNU injection and, (iv) bone marrow cells rapidly underwent apoptosis following MNU treatment, demonstrating that PARP is a survival factor playing an essential and positive role during DNA damage recovery and survival.
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  • 41
    ISSN: 1573-4919
    Keywords: PARP ; poly(ADP-ribosyl)ation ; apoptosis ; DNA replication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We have focused on the roles of PARP and poly(ADP-ribosyl)ation early in apoptosis, as well as during the early stages of differentiation-linked DNA replication. In both nuclear processes, a transient burst of PAR synthesis and PARP expression occurs early, prior to internucleosomal DNA cleavage before commitment to apoptosis as well as at the round of DNA replication prior to the onset of terminal differentiation. In intact human osteosarcoma cells undergoing spontaneous apoptosis, both PARP and PAR decreased after this early peak, concomitant with the inactivation and cleavage of PARP by caspase-3 and the onset of substantial DNA and nuclear fragmentation. Whereas 3T3-L1, osteosarcoma cells, and immortalized PARP +/+ fibroblasts exhibited this early burst of PAR synthesis during Fas-mediated apoptosis, neither PARP-depleted 3T3-L1 PARP-antisense cells nor PARP -/- fibroblasts showed this response. Consequently, whereas control cells progressed into apoptosis, as indicated by induction of caspase-3-like PARP-cleavage activity, PARP-antisense cells and PARP -/- fibroblasts did not, indicating a requirement for PARP and poly(ADP-ribosyl)ation of nuclear proteins at an early reversible stage of apoptosis. In parallel experiments, a transient increase in PARP expression and activity were also noted in 3T3-L1 preadipocytes 24 h after induction of differentiation, a stage at which ~95% of the cells were in S-phase, but not in PARP-depleted antisense cells, which were consequently unable to complete the round of DNA replication required for differentiation. PARP, a component of the multiprotein DNA replication complex (MRC) that catalyzes viral DNA replication in vitro, poly(ADP-ribosyl)ates 15 of ~40 MRC proteins, including DNA pol α, DNA topo I, and PCNA. Depletion of endogenous PARP by antisense RNA expression in 3T3-L1 cells results in MRCs devoid of any DNA pol α and DNA pol δ activities. Surprisingly, there was no new expression of PCNA and DNA pol α, as well as the transcription factor E2F-1 in PARP-antisense cells during entry into S-phase, suggesting that PARP may play a role in the expression of these proteins, perhaps by interacting with a site in the promoters for these genes.
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  • 42
    ISSN: 1573-4919
    Keywords: breast cancer cells ; anti-apoptotic genes ; apoptosis ; progesterone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Progesterone inhibits the proliferation of normal breast epithelial cells in vivo, as well as breast cancer cells in vitro. But the biologic mechanism of this inhibition remains to be determined. We explored the possibility that an antiproliferative activity of progesterone in breast cancer cell lines is due to its ability to induce apoptosis. Since p53, bcl-2 and survivin genetically control the apoptotic process, we investigated whether or not these genes could be involved in the progesterone-induced apoptosis. We found a maximal 90% inhibition of cell proliferation with T47-D breast cancer cells after exposure to 10 μM progesterone for 72 h. Control progesterone receptor negative MDA-231 cancer cells were unresponsive to 10 μM progesterone. The earliest sign of apoptosis is translocation of phosphatidylserine from the inner to the outer leaflet of the plasma membrane and can be monitored by the calcium-dependent binding of annexin V in conjunction with flow cytometry. After 24 h of exposure to 10 μM progesterone, cytofluorometric analysis of T47-D breast cancer cells indicated 43% were annexin V-positive and had undergone apoptosis and no cells showed signs of cellular necrosis (propidium iodide negative). After 72 h of exposure to 10 μM progesterone, 48% of the cells had undergone apoptosis and 40% were annexin V positive/propidium iodide positive indicating signs of necrosis. Control untreated cancer cells did not undergo apoptosis. Evidence proving apoptosis was also demonstrated by fragmentation of nuclear DNA into multiples of oligonucleosomal fragments. After 24 h of exposure of T47-D cells to either 1 or 10 μM progesterone, we observed a marked down-regulation of protooncogene bcl-2 protein and mRNA levels. mRNA levels of survivin and the metastatic variant CD44 v7-v10 were also downregulated. Progesterone increased p53 mRNA levels. These results demonstrate that progesterone at relative high physiological concentrations, but comparable to those seen in plasma during the third trimester of human pregnancy, exhibited a strong antiproliferative effect on breast cancer cells and induced apoptosis.
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  • 43
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    Molecular and cellular biochemistry 200 (1999), S. 51-57 
    ISSN: 1573-4919
    Keywords: smokeless tobacco ; apoptosis ; nitric oxide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Smokeless tobacco usage is, a growing public health concern in the United States. Lesions of the oral cavity have been clearly linked to smokeless tobacco use. The objective of this study was to determine the biochemical effects of smokeless tobacco extract (STE) exposure upon hamster cheek pouch cell (HCPC-1) cultures. HCPC-1 cells were exposed to a 5 -fold dose-range of STE (0.5, 1.0 and 2.5%) over a time-course of 24-96 h. Following each exposure we measured various biochemical parameters of cell proliferation and cell death. Cell viability, cell cycle progression and S-phase DNA synthesis were measured as markers of cell proliferation. We measured lactate dehydrogenase leakage as a marker of cell membrane damage and cell death due to necrosis. No significant alterations were observed in cell cycle progression and cell proliferation as a result of exposure to STE. LDH measured colorimetrically indicated no significant effect with the lower doses (0.5, 1.0 and 2.5% STE). Apoptosis measured as the A0 peak and by the TUNEL procedure revealed that STE caused significant rates of apoptosis. Maximal apoptosis was noted between 48-96 h. In order to probe the mechanism further we measured the levels of nitrites as an indicator of nitric oxide (NO) in the media. NO levels were significantly elevated at the doses that caused an induction of apoptosis. The results from this study indicate that STE causes a dose-dependent induction of apoptosis and that this is mediated by nitric oxide.
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  • 44
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    Molecular and cellular biochemistry 193 (1999), S. 37-42 
    ISSN: 1573-4919
    Keywords: Rho ; GTPase ; toxins ; Clostridium ; signal transduction ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The Rho family small GTPases are members of the Ras superfamily of small GTPases. Rho proteins were first determined to act as key regulators of many types of actin cytoskeletal-dependent cellular functions. Recent work by several investigators indicates that Rho GTPases are also critical modulators of several important intracellular and nuclear signal transduction pathways. Certain clostridial toxins and exoenzymes covalently modify, and thereby inactivate, specific types of Rho family GTPases. As such, these microbial enzymes have proven invaluable in helping to identify structural and functional attributes of Rho GTPases.
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  • 45
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    Molecular and cellular biochemistry 193 (1999), S. 103-108 
    ISSN: 1573-4919
    Keywords: Poly(ADP-ribose) polymerase ; Drosophila melanogaster ; alternative splicing ; apoptosis ; DNA repair ; development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Poly(ADP-ribose) polymerase (PARP) is conserved in eukaryotes. To analyze the function of PARP, we isolated and characterized the gene for PARP in Drosophila melanogaster. The PARP gene consisted of six translatable exons and spanned more than 50 kb. The DNA binding domain is encoded by exons 1-4. Although the consensus cleavage site of CED-3 like protease during apoptosis is conserved from human to Xenopus laevis PARPs, it is neither conserved in the corresponding region of Drosophila nor Sarcophaga peregrina. There are two cDNAs species in Drosophila. One cDNA could encode the full length PARP protein (PARP I), while the other is a truncated cDNA which could encode a partial-length PARP protein (PARP II), which lacks the automodification domain and is possibly produced by alternative splicing. The expression of these two forms of PARP in E. coli demonstrated that while PARP II has the catalytic NAD-binding domain and DNA-binding domain it is enzymatically inactive. On the other hand PARP I is active. A deletion mutant of PARP gene could grow to the end of embryogenesis but did not grow to the adult fly. These results suggest that the PARP gene plays an important function during the development of Drosophila.
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  • 46
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    Molecular and cellular biochemistry 193 (1999), S. 119-125 
    ISSN: 1573-4919
    Keywords: benzamides ; nicotinamides ; apoptosis ; inflammation ; NF-kB ; DNA repair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Our laboratory has concentrated on the possible regulation the benzamides and nicotinamides may have on the processes of DNA repair and apoptosis. Recent reports [14-16] have suggested that both apoptosis and inflammation are regulated by the transcription factor NF-kB. We have initiated studies regarding the hypothesis that the benzamides and nicotinamides could inhibit the production of tumor necrosis factor alpha (TNFalpha) and the inflammatory response as well as induce apoptosis via inhibition of NF-kB. Our data have shown that nicotinamide and two N-substituted benzamides, metoclopramide (MCA) and 3-chloroprocainamide (3-CPA), gave dose dependent inhibition of lipopolysacharide induced TNFalpha in the mouse within the dose range of 10-500 mg/kg. Moreover, lung edema was prevented in the rat by 3 ï 50 mg/kg doses of 3-CPA or MCA, and 100-200 μM doses of MCA could also inhibit NF-kB in Hela cells. Taken together these data strongly support the notion that benzamides and nicotinamides have potent anti-inflammatory and antitumor properties, because their primary mechanism of action is regulated by inhibition at the gene transcription level of NF-kB, which in turn inhibits TNFalpha and induces apoptosis.
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  • 47
    ISSN: 1573-4919
    Keywords: antisense oligonucleotide ; apoptosis ; cAMP-dependent protein kinase ; cancer cells ; growth inhibition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The enhanced expression of the RIα subunit of cyclic AMP-dependent protein kinase type 1 (PKA-I) has been correlated with cancer cell growth. We have investigated the effects of sequence-specific inhibition of RIα gene expression on the growth of MCF-7 human breast cancer cells. We report that RIα antisense treatment results in a reduction in RIα expression at both mRNA and protein levels and inhibition of cell growth. The growth inhibition was accompanied by changes in cell morphology, cleavage of poly(ADP-ribose) polymerase (PARP) and appearance of apoptotic nuclei. In addition, bcl-2 protein level was reduced and p53 expression increased in growth arrested cells. Interestingly, RIα antisense inhibited cell viability and induced apoptosis in the absence of p53, suggesting that these actions of RIα antisense are exerted independent of p53. In contrast, two- and four-base mismatched control oligonucleotides had no effect on either cell growth or morphology. These results demonstrate that the RIα antisense, which efficiently depletes the growth stimulatory molecule RIα, induces cell differentiation and apoptosis, providing a new approach to combat breast cancer cell growth.
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  • 48
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    Molecular and cellular biochemistry 197 (1999), S. 97-108 
    ISSN: 1573-4919
    Keywords: neutrophil ; PKC ; TNF-α ; apoptosis ; DNA fragmentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract In the present study we investigated the TNF-α induced signal transduction mechanism in human neutrophil. Exogenously added TNF-α affects both PKC activity and its translocation from cytosol to the membrane. Endogenous protein phosphorylation pattern is inhibited in TNF-α induced neutrophil in Ca-dependent and Ca-independent manner, including a major 47 and 66 kDa cytosolic proteins, which may be implicated in superoxide anion generation. However TNF-α dose dependently enhances the expression of ζ-PKC isotype but not the β-PKC. Morphology and cell cytotoxicity are studied in TNF-α treated neutrophil to understand the TNF-α induced cell death or apoptosis and these experiment is further confirmed by DNA fragmentation analysis. These results clearly demonstrate that TNF-α induces cellular death of human neutrophil at least in part by enhanced expression of Ca-independent ζ-PKC. These observations provide an insight towards understanding the function of ζ-PKC in apoptotic pathway.
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  • 49
    ISSN: 1573-4919
    Keywords: rotenone ; apoptosis ; oncogenes ; liver cancer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Rotenone decreases the incidence of hepatocellular carcinoma and lowers rates of hepatocellular proliferation. In an effort to delineate mechanisms involved, the in vivo effect of rotenone on liver mitochondrial metabolism, apoptotic machinery as well as elements of the hepatic signal transduction pathways were investigated. Mitochondria from livers of male B6C3F1 mice fed a standard diet containing 600 ppm rotenone for 7 days were uncoupled or inhibited when succinate or glutamate plus malate were used as the substrate, respectively. These livers also showed a significant increase in apoptosis compared with control livers. Furthermore, rotenone increased the expression of c-myc mRNA to 5-fold of control values within 3 days, an effect which was still observed (3-fold) after 7 days. Levels of p53 mRNA were also increased 3-fold after 1 day, but declined to control levels by 7 days. Rotenone also caused a transient, yet marked increase in liver particulate glyceraldehyde phosphate dehydrogenase (GAPDH) protein expression, while it did not alter the expression of the cytosolic form of the enzyme. Conversely, mRNA of the proto-oncogene H-ras showed a decline of 35% after 3 days of rotenone treatment, and remained diminished for the duration of the experiment. These data suggest that rotenone may act as an anticancer agent by diminishing mitochondrial bioenergetics which prevents basal hepatocyte proliferation and lowers the threshold for liver cells with DNA damage to undergo apoptosis.
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  • 50
    ISSN: 1573-4943
    Keywords: Two-dimensional electrophoresis ; MALDI-MS ; apoptosis ; RNA polymerase B transcription factor 3
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Apoptosis or programmed cell death is essential in the process of controlling lymphocyte growth and selection. We identified RNA polymerase B transcription factor 3 (BTF3), which is associated with anti-IgM antibody-mediated apoptosis, using a subclone of the human Burkitt lymphoma cell line BL60. To identify the transcription factor BTF3, which is expressed only in minor amounts, we used preparative high-resolution two-dimensional gel electrophoresis (2DE) employing carrier ampholytes for isoelectric focusing. Comparison of the 2DE protein patterns from apoptotic and nonapoptotic cells showed BTF3 as a predominantly altered protein spot. The characterization of the differentially expressed transcription factor and 13 marker proteins described in this study were performed by internal Edman microsequencing and/or by peptide mass fingerprinting using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The proteome analysis was significantly improved by performing the newly developed preparative high-resolution two-dimensional gels employing high protein concentrations.
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  • 51
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    Investigational new drugs 17 (1999), S. 227-240 
    ISSN: 1573-0646
    Keywords: apoptosis ; protein kinase C ; sphingoid bases ; safingol ; diglyceride ; bryostatin 1 ; staurosporine ; 7-hydroxy staurosporine (UCN-01) ; 4′-N-benzoyl staurosporine (CGP-41251) ; calphostin C (UCN-1028c)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Neoplastic cell survival is governed by a balance between pro-apoptotic and anti-apoptotic signals. Noteworthy among several anti-apoptotic signaling elements is the protein kinase C (PKC) isoenzyme family, which mediates a central cytoprotective effect in the regulation of cell survival. Activation of PKC, and subsequent recruitment of numerous downstream elements such as the mitogen-activated protein kinase (MAPK) cascade, opposes initiation of the apoptotic cell death program by diverse cytotoxic stimuli. The understanding that the lethal actions of numerous antineoplastic agents are, in many instances, antagonized by cytoprotective signaling systems has been an important stimulus for the development of novel antineoplastic strategies. In this regard, inhibition of PKC, which has been shown to initiate apoptosis in a variety of malignant cell types, has recently been the focus of intense interest. Furthermore, there is accumulating evidence that selective targeting of PKC may prove useful in improving the therapeutic efficacy of established antineoplastic agents. Such chemosensitizing strategies can involve either (a) direct inhibition of PKC (e.g., following acute treatment with relatively specific inhibitors such as the synthetic sphingoid base analog safingol, or the novel staurosporine derivatives UCN-01 and CGP-41251) or (b) down-regulation (e.g., following chronic treatment with the non-tumor-promoting PKC activator bryostatin 1). In preclinical model systems, suppression of the cytoprotective function(s) of PKC potentiates the activity of cytotoxic agents (e.g., cytarabine) as well as ionizing radiation, and efforts to translate these findings into the clinical arena in humans are currently underway. Although the PKC-driven cytoprotective signaling systems affected by these treatments have not been definitively characterized, interference with PKC activity has been associated with loss of the mitogen-activated protein kinase (MAPK) response. Accordingly, recent pre-clinical studies have demonstrated that pharmacological disruption of the primary MEK-ERK module can mimic the chemopotentiating and radiopotentiating actions of PKC inhibition and/or down-regulation.
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  • 52
    ISSN: 1573-0646
    Keywords: UCN-01 ; IL-2 receptor ; Fas ; Fas-ligand ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract 7-hydroxystaurosporine (UCN-01) is a new anticancer agentwhich exerts an inhibitory effect on cell cycle check points andis currently under phase I clinical trials in US and Japan.Preliminary clinical data indicated that UCN-01 remained inplasma at high concentrations for long periods of time. Thisunavoidable high plasma drug exposure is likely to lead tohematological toxicities in patients. In the present study,cultured human peripheral blood lymphocytes (PBLs) were used toevaluate the possible hematological toxicities of UCN-01treatment. UCN-01 induces apoptosis, and the induction ofapoptosis-related surface markers were also examined toinvestigate the involvement of these molecules in UCN-01-inducedapoptosis in PBLs. in vitroviability of PBLs wasdecreased by high dose of UCN-01 (25 μM, 3-day exposure). Thiseffect of UCN-01 was significantly suppressed by the presence ofhuman serum, suggesting that some specific inhibitory factor(s)in human serum may antagonize the lympholytic effect of UCN-01.The percentage of annexin V-positive PI-negative cells increasedwith exposure to UCN-01 in a time- and dose-dependent manner; byup to 30.3% after exposure to 25 μM UCN-01 for 3 days.At the same time, the expression of both interleukin-2 receptor(IL-2R, CD25) and Fas (CD95), analyzed by flow cytometry, wasinduced. Con A-stimulated PBLs were more sensitive toUCN-01-induced apoptosis than non-stimulated lymphocytes andUCN-01 increased the sFas-L released into culture medium from conA-stimulated PBLs. Therefore, lymphocyte depletion mediated byactivation-induced apoptosis is likely to occur in patientstreated with UCN-01 at high doses.
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  • 53
    ISSN: 1573-0778
    Keywords: apoptosis ; Bcl-2 ; fixed-bed ; hollow fibre ; hybridoma ; perfusion ; protein-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Apoptosis is an active, genetically determined death mechanism which can be induced by a wide range of physiological factors and by mild stress. It is the predominant form of cell death during the production of antibodies from murine hybridoma cell lines. A number of studies have now demonstrated that the suppression of this death pathway, by means of over-expression of survival genes such as bcl-2, results in improved cellular robustness and antibody productivity during batch culture. In the present study, the influence of bcl-2 expression on hybridoma productivity in two high density perfusion bioreactor systems was investigated. In the first system, a fixed-bed reactor, the DNA content in the spent medium was 25% higher in the control (TB/C3-pEF) culture than that found in the bcl-2 transfected (TB/C3-bcl2) cultures at all perfusion rates. This is indicative of a higher level of cell death in the control cell line. The average antibody concentration for the TB/C3-pEF cell line was 14.9 mg L-1 at perfusion rates of 2.6 and 5.2 d-1. However, for the TB/C3-bcl2 cell line it was 33 mg L-1 at dilution rates of 2 and 4 d-1. A substantial increase in antibody concentration was also found in the Integra Tecnomouse hollow fibre reactor. The antibody titre in the TB/C3-bcl2 cassette was nearly 100% higher than that in the TB/C3-pEF cassette during the cultivation period which lasted 6 weeks. Clearly, these results demonstrate the positive impact of bcl-2 over-expression on production of antibody in hybridoma perfusion cultures.
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  • 54
    ISSN: 1573-0778
    Keywords: antibody productivity ; apoptosis ; BAG-1 ; Bcl-2 ; cell survival ; hybridoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Human bcl-2 and bag-1 DNA were introduced into mouse hybridoma 2E3- O cells and expressed. The expression of bcl-2 in BCMGneo-bcl2 transfectants was confirmed by ELISA and that of bag-1 in pZeo-bag1 was confirmed by western blotting. In batch cultures, the over-expression of bcl-2 prolonged the culture period by 2 days and co-expression of bcl-2 and bag-1 prolonged the culture period by 3 days. The delayed increase in the dead cell number in culture of the bcl-2 and bag-1 cotransfectant indicated the additional antiapoptosis effect of bcl-2 and bag-1 cotransfection in comparison with the bcl-2 only transfection. The bcl-2 transfectants (2E3O-Bcl2) produced antibody twofold per batch culture in comparison with 2E3-O cells transfected with BCMGSneo (2E3O-Mock). Enhancement of this MoAb production was due to the improved survival of the cells and was not due to stimulation of antibody production rate per cell by Bcl-2 expression. And the bcl-2 and bag-1 co-transfectant (2E3O-Bcl2-BAG1) produced antibody approximately fourfold of 2E3O-Mock per batch culture. Enhancement of this MoAb production was due to the improved survival of the cells and was partly due to stimulation of MoAb production rate per cell in the non-growing phase by the cotransfection. The method to engineer hybridoma cells genetically with bcl-2 and bag-1 for increasing viability and productivity would be widely applied for improving antibody productivity of hybridoma cultures.
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  • 55
    ISSN: 1573-0778
    Keywords: annexin V ; Apo-2 ligand ; apoptosis ; Cytostar-T® scintillating microplates ; flow cytometry ; lymphotoxin (LT)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The translocation of phospholipids across the plasma membrane has been widely documented as one of the earliest measurable biochemical events of apoptosis. Using fluorescently labelled annexin V, which preferentially binds phosphatidylserine (PS) in the presence of Ca2+, the externalization of PS can be measured and apoptosis quantified using flow cytometry. Conventional detection methods utilizing annexin V, while faster than in situ DNA end-labelling or DNA laddering, require extensive sample preparation which may compromise samples and makes rapid, high volume screening prohibitive. This paper describes a novel assay for the measurement of apoptosis based upon binding of radiolabelled annexin V to apoptotic cells attached to the growth surface of a 96-well scintillating microplate (Cytostar-T®). We compared measurements of apoptosis made by flow cytometry to those obtained with the scintillating microplate in three model systems, treatment of: mouse connective tissue (L-M) cells with lymphotoxin (LT), human lung carcinoma (H460) cells with Apo-2 ligand and human umbilical vein endothelial (HUVE) cells with staurosporine. In this assay, we compare both direct and indirect labelling methods by utilizing either iodinated annexin V or biotinylated annexin V/[35S] streptavidin to radiolabel apoptotic cells. The signal detected is a direct consequence of the binding of annexin V to externalized PS on apoptotic cells and the proximity of the label to the base of the plate. Using this method, separation of bound and unbound radiolabel signal occurs directly within the well resulting in a sensitive assay that requires minimal manipulation and can accomodate a large number of samples.
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  • 56
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    Cardiovascular drugs and therapy 13 (1999), S. 289-294 
    ISSN: 1573-7241
    Keywords: heart failure ; apoptosis ; protein kinases ; caspases ; DNA damage ; cardiomyocytes ; β-blocks
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Apoptosis as defined by contemporary science describes a form of cell death that involves discrete genetic and molecular programs, de novo protein expression and unique cellular phenotype. Evidence for the existence of apoptosis in the human heart has been reported in various cardiac diseases, including ischemic and non-ischemic heart failure, myocardial infarction and arrhythmias. Among the most potent stimuli that elicit cardiomyocyte apoptosis are: oxygen radicals (including NO), cytokines, (FAS/TNFα family of cytokines) and growth factors/energy deprivation. Several complex signal transduction pathways have been implicated in execution of cardiomyocyte apoptosis, including: Fas/TNFα receptors signaling, stress or mitogen activated protein kinases (SAPK/MAPK), sphingolipids metabolites (ceramide), G-protein coupled receptor (GPCR) signaling (Gαi, Gαq) and NFkB activation. Apoptosis of cardiac myocytes may contribute to progressive pump-failure, arrhythmias and cardiac remodeling. The recognition of numerous molecular targets associated with cardiomyocyte apoptosis that are amenable for pharmacologic manipulation, may provide novel therapeutic strategies for diverse cardiac ailments, as recently suggested by pharmacologic studies in experimental animals.
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  • 57
    ISSN: 1573-7373
    Keywords: apoptosis ; cell death ; C6 glioma ; nerve growth factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Nerve growth factor (NGF) plays an important physiological role in differentiation and survival of various types of neurons. Glial cells and glial tumor cells synthesize multiple neurotrophic factors including NGF and secrete them into the surrounding environment; however, the mechanisms of NGF and the significance of NGF receptors have not been studied in detail. The C6 glioma cell line can synthesize NGF, respond to exogenous application of NGF and stimulate the expression of NGF receptor in an autocrine manner. In order to determine the significance of such an NGF autocrine system, the effects of exposure to exogenous NGF and deprivation of endogenous NGF were examined in a C6 glioma cell line in vitro. Exogenous NGF significantly inhibited maintenance of the cell number and thymidine incorporation. Morphological changes, including the formation of growth cones, outgrowth of processes and cellular hypertrophy, were observed, concurrently, indicating that exogenous NGF stimulated differentiation and thereby inhibited proliferation of the cells. Deprivation of endogenous NGF with anti-NGF antibody elicited a rapid decrease in cell number and thymidine incorporation, and led almost all of the cells to death within 8 days. The protein synthesis inhibitor, cycloheximide, strongly inhibited the death of NGF-deprived cells, suggesting the involvement of an active process requiring synthesis of suicide proteins. These findings imply that the NGF autocrine system plays a significant role in regulating the differentiation and survival of C6 glioma cells, similarly to neuronal cells.
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  • 58
    ISSN: 1573-7373
    Keywords: glutamate ; glioblastoma ; apoptosis ; Bcl-2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Glutamate has been shown to function as a toxic agent in neuronal and glial cells, as well as an excitatory neurotransmitter throughout the central nervous system. In the present study, we examined the effect of increasing glutamate concentration on the induction of apoptosis in the two human glioblastoma cell lines GB-4 and GB-12. Glutamate exposure caused cell death of GB-4 and GB-12 in a dose-dependent manner. The cells were found to die via apoptosis in response to glutamate based on the following criteria: propidium iodide (PI) staining, H–E staining, electron microscopic analysis, and the TdT-mediated dUTP-biotin nick end labeling (TUNEL) method. The glutamate-induced apoptosis appears to involve the modulation of Bcl-2 family gene products such as Bcl-2, Bcl-xL, and Bax-α. Both Bcl-2 and Bcl-xL were down-regulated by glutamate at 24 h and further at 48 h. The apoptosis-promoting product p21 Bax-α was also down-regulated in GB-12 but slightly up-regulated in GB-4, accompanied by generation of variant form of p18 Bax-α in both cell lines. These findings suggest that glutamate toxicity results in cellular death via an apoptotic mechanism which appears to involve the Bcl-2/Bax-α molecular complex.
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  • 59
    ISSN: 1573-7365
    Keywords: Cerebral ischemia ; focal ischemia ; mutant mice ; bcl-2 ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Neuronal death after brain ischemia is mainly due to necrosis but there is also evidence for involvement of apoptosis. To test the importance of apoptosis, we investigated the effect of targeted disruption of the apoptosis-suppressive gene bcl-2 on the severity of ischemic brain injury. Transient focal ischemia for 1 hour was induced by occlusion of the middle cerebral artery in homozygous (n=7) and heterozygous (n = 6) bcl-2 knockout mice as well as in their wildtype littermates (n=5). Bcl-2 ablation did not influence cerebral blood flow but it significantly increased infarct size and neurological deficit score at 1 day after reperfusion in a gene-dose dependent manner. The exacerbation of tissue damage in the absence of Bcl-2 underscores the importance of apoptotic pathways for the manifestation of ischemic injury after transient vascular occlusion.
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  • 60
    ISSN: 1573-7373
    Keywords: glioma ; immunohistochemistry ; MIB-1 ; apoptosis ; TUNEL
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract With the introduction of new (immuno-)histochemical techniques it is now possible to assess rates of proliferation and apoptosis in brain gliomas using archival paraffin embedded material. As proliferation and apoptosis are related to tumour growth rate quantification of these processes has prognostic value and is related to tumour grading. In this study we assessed the proliferation rate by measuring the Ki-67 labelling index using the MIB-1 antibody (MIB-LI) and the apoptotic rate using the in situ labelling of DNA strand breaks with TUNEL (TUNEL-LI) in 315 supratentorial gliomas. MIB-LI and TUNEL-LI in astrocytomas (A) where significantly lower compared to anaplastic astrocytomas (AA), glioblastomas (GBM) and oligodendroglial tumours [oligodendrogliomas (O) and anaplastic oligodendrogliomas (AO)]. MIB-LI and TUNEL-LI were significantly lower in AA compared to GBM. In astrocytic tumours MIB-LI and TUNEL-LI appeared to be correlated. As the distinction between A and AA is of clinical value but can be difficult histomorphologically we analysed the prognostic value of MIB-LI and TUNEL-LI in gliomas with particular emphasis on A and AA. MIB-LI below 10% was of prognostic value in A and AA, O and AO but not in GBM on univariate survival analysis. TUNEL-LI was of no prognostic value. With multivariate survival analysis MIB-LI lost prognostic significance in O and AO. Astrocytomas with a gemistocytic component (AG) are similar to A with respect to survival and MIB-LI and TUNEL-LI. MIB-LI is of independent prognostic value in A and AA. Assessment of MIB-LI in A and AA can be used as an aid in distinguishing A and AA.
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  • 61
    ISSN: 1573-7373
    Keywords: apoptosis ; astrocytoma ; glioma ; growth inhibition ; protein kinase C ; signal transduction ; UCN-01
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Recent studies in our laboratory have shown that UCN-01 (7-hydroxystaurosporine), which is a derivative of the non-selective protein kinase inhibitor staurosporine that exhibits relative selectivity for protein kinase C (PKC), is a potent inhibitor of glioma growth in in vitro and in vivo models. This agent exhibits both cytotoxic and cytostatic effects, depending on the time period of drug exposure. In the present study, we examined whether UCN-01-induced cytotoxicity correlated with the induction of apoptosis, and characterized further the time course of this process as a prelude to application of UCN-01 in clinical trials. We first demonstrated that the cytotoxic effects of UCN-01 were associated with the induction of morphological features of apoptosis. Secondly. we identified electrophoretic features of apoptosis semiquantitatively at a series of time points using field inversion gel electrophoresis. These studies showed a peak in the induction of high-molecular-weight DNA fragmentation after 3–6 days of drug treatment. Thirdly, we measured the percentage of cells undergoing apoptosis at various time points using a terminal transferase-catalyzed in situ end-labeling technique, which confirmed a time- and concentration-dependent increase in apoptotic cell numbers. This correlated with a progressive decrease in the percentage of cells that were viable as assessed by trypan blue exclusion. Cell killing peaked within 2–4 days after beginning UCN-01 treatment, but continued at a lower level in the ensuing days. Taken together, these studies demonstrated that extended periods of exposure to UCN-01 are needed for optimal manifestation of cytotoxic effects against glioma cells, a factor that must be taken into consideration in the design of future clinical trials with this agent for malignant gliomas.
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  • 62
    ISSN: 1573-7373
    Keywords: germinoma ; spontaneous regression ; multiple intracranial ; germ cell tumor ; immunological mechanism ; apoptosis ; MIB-1 index
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A case of multiple intracranial germ cell tumor in which a pineal tumor regressed spontaneously after resection of the cerebellar mass is reported. Immunohistochemical staining of the cerebellar mass showed that most of the infiltrating lymphocytes were positive for CD3 and CD8. The anti-Ki-67 monoclonal antibody MIB-1 staining of the resected tumor revealed a high MIB-1 positivity ratio (36.1%) among the large tumor cells, and TUNEL staining demonstrated that positivity in up to 6% of the tumor cells. Possible mechanisms responsible for this spontaneous regression including immunological responses and apoptosis induced by T lymphocytes are discussed.
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  • 63
    ISSN: 1573-7373
    Keywords: apoptosis ; cell cycle ; cell death ; medulloblastoma ; T cells ; tumour necrosis factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We studied the effect of the treatment of a medulloblastoma cell line by human T cells derived soluble factors. Medulloblastoma is one of the more common aggressive solid neoplasms in children for which there is no adequate therapy. Cell lines established from such tumours may be helpful to test the effect of various molecules on cell proliferation. Previous studies have suggested that T cell-derived factors may be toxic for the medulloblastoma cell line Dev. Cytokines were thought to mediate this effect. In this paper, we described changes in morphology, survival and cell cycle induced in Dev cells cocultured with human T cell lines chronically infected with a retrovirus (HTLV-I) and known to secrete high level of cytokines TNFα, IL1α and IL6. Such cocultures resulted in the death of a part of Dev cells and in decreased proliferation of surviving cells, associated with morphological changes and increase in vimentin expression. Treatment with conditioned medium from infected Dev cells, containing virus induced cytokines, triggered the same effect. Reduction of these effects by TNFα deprivation of conditioned medium suggested that this cytokine may be implicated. Direct treatment of Dev cells with recombinant cytokines indicated that TNFα, but not IL1 or IL6, is associated with Dev cell alterations. TNFα was shown to induce the death of Dev cells by an apoptotic pathway. Furthermore, TNFα had a bimodal effect on the cell cycle of surviving Dev cells. These differential effects of such cytokines on medulloblastoma cells could be therefore of interest for immunotherapy of these tumours.
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  • 64
    ISSN: 1573-7373
    Keywords: adenoviral gene transfer ; p53 ; apoptosis ; experimental malignant gliomas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Adenoviral-mediated gene transfer for the treatment of experimental intrinsic malignant brain neoplasms holds promise. The role, however, of intracellular, adenoviral-mediated p53 expression to inhibit growth of experimental human intracranial malignant gliomas remains largely unexplored. Using the AdCMV.p53 vector we measured the in vitro expression of p53 and the resultant effect upon U251 human malignant glioma cellular proliferation. We further measured the survival of nude mice after intracranial injection of the infected vs. control U251 cells. The growth of the infected U251 cells was inhibited when compared to both the uninfected cells and cells infected with the control vector (AdCMV.Null). Agarose gel electrophoresis confirmed the AdCMV.p53-dependent cellular apoptosis. Nude mice having intracranial injections of the U251 cells infected with the control (AdCMV.Null) vector showed diminished survival. In contrast, mice having intracranial injections of the cells infected with the AdCMV.p53 vector showed 100% survivorship measured 100 days after treatment. Gene therapy via the AdCMV.p53 viral vector holds promise for the clinical treatment of human malignant gliomas.
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  • 65
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    Journal of neuro-oncology 41 (1999), S. 159-166 
    ISSN: 1573-7373
    Keywords: neuroblastoma ; chemotherapy ; apoptosis ; dopamine receptors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract While neuroscientists are often involved in the assessment and care of patients with central nervous system tumors, they are only rarely involved in the case of peripheral nervous system neoplasia. Neuroblastoma is a childhood tumor of the primitive sympathetic nervous system. It is at once one of the most common and one of the most deadly tumors of childhood. The prognosis for children with this tumor has not changed in the past two decades. Clearly, a fresh approach to neuroblastoma is needed. The neuroscientist has much to add to our understanding and treatment of neuroblastoma and its sequelae. Conversely, neuroblastoma has much to teach us regarding the normal development of the neural crest and the aberrant loss of neurons in this lineage. A neuroscientist's approach to neuroblastoma, its biology and clinical features, is presented herein.
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  • 66
    ISSN: 1573-7373
    Keywords: apoptosis ; chemotherapy ; human cell lines ; lovastatin ; medulloblastoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Medulloblastoma is a malignant paediatric central nervous system tumor with a poor prognosis, stimulating the evaluation of improved treatment strategies. Lovastatin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, is currently used to treat patients with hypercholesterolemia. This compound also inhibits the production of non-steroidal mevalonate derivatives that are implicated in the control of cellular proliferation, and can induce cell-cycle arrest in vitro. We recently showed that lovastatin inhibited growth and promoted apoptosis of neuroblastoma, the peripheral nervous system ‘cousin’ of medulloblastoma. Therefore the potential of lovastatin as a possible anticancer drug against medulloblastoma was evaluated in vitro. Four medulloblastoma cell lines, Daoy, UW228, D341 Med and D283 Med, were treated with 1–40 µM of lovastatin in vitro. Analysis of cell morphologic changes, cell viability, DNA fragmentation and flow cytometry in all four cell lines showed growth inhibition and induction of apoptosis with lovastatin treatment. As little as 10 µM of lovastatin was sufficient to cause a marked reduction in cell numbers, and more than 20 µM of lovastatin induced 〉90% cells to undergo apoptosis, after intervals ranging between 36 and 96 h, depending on the cell line. Lovastatin induced apoptosis in these cell lines was concomitant with cell cycle arrest in G1. The attached cell lines UW228 and Daoy were more sensitive to lovastatin than D283 Med and D341 Med. Daoy cells which survived several cycles of lovastatin treatment could still be induced to undergo apoptosis after longer treatment times. The efficient induction of apoptosis by lovastatin favours this drug as a potential new avenue of therapeutic intervention for medulloablastoma.
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  • 67
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    Journal of neuro-oncology 43 (1999), S. 237-239 
    ISSN: 1573-7373
    Keywords: primary CNS lymphoma ; glucocorticoids ; cerebral edema ; apoptosis ; bcl-2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Glucocorticoid therapy may result in the rapid resolution of cerebral mass lesions in patients with primary CNS lymphoma. Since glucocorticoids will obscure the histological diagnosis of primary CNS lymphoma upon biopsy, steroids should be withheld if primary CNS lymphoma is a likely diagnosis by neuroradiological criteria. The lympholytic effect of glucocorticoids is mediated by cytoplasmic steroid receptors which are translocated to the nucleus and signal apoptosis. Glucocorticoid-induced apoptosis of lymphoid cells does not require wild-type p53 activity, seems not to depend on caspase activation, but is attenuated by the bcl-2 protooncogene product. Long-term glucocorticoid therapy of primary CNS lymphoma is not recommended because relapse is probably inevitable and because of the prominent side effects of long-term glucocorticoid treatment. Further, long-term glucocorticoid treatment is contraindicated in immunocompromised patients with primary CNS lymphoma.
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  • 68
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    Journal of neurocytology 28 (1999), S. 149-159 
    ISSN: 1573-7381
    Keywords: prion disease ; transmissible spongiform encephalopathy ; cell culture ; toxicity ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The aetiological agent of prion disease is proposed to be an aberrant isoform of the cell surface glycoprotein known as the prion protein (PrPc). This pathological isoform (PrPSc) is abnormally deposited in the extracellular space of diseased CNS. Neurodegeneration in these disease has been shown to be associated with accumulation of PrPSc in affected tissue. To investigate the possible uptake mechanisms that may be required for PrPSc-induced neurodegeneration we studied the cellular trafficking of the neurotoxic fragment, PrP106-126. We were able to detect, by fluorescence microscopy, PrP106-126 inclusions in murine neurones, astrocytes and microglia in vitro. These inclusions were abundant after 24 hour exposure and still present 48h post-exposure. Shorter exposure times yielded only occasional cells with inclusions. Large extracellular aggregates of PrP106-126 could also be detected, which appeared in a time dependent manner. The appearance of inclusions or aggregates was not dependent on PrPc expression as determined by exposure of peptides from PrP-null mice. Using transmission electron microscopy and gold particle detection, positively labelled osmiophilic inclusions of peptide could be detected in the cytoplasm of exposed cells. These results demonstrate that cultured cells are capable of sequestering PrP106-126 and may indicate uptake pathways for PrPSc in various cell types. Toxicity of PrP106-126 may thus be mediated via a sequestration pathway that is not effective for this peptide in PrP-null cells.
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  • 69
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    Bioscience reports 19 (1999), S. 345-354 
    ISSN: 1573-4935
    Keywords: Glycolipid ; apoptosis ; intracellular traffic ; multidrug resistance ; ovarian carcinoma ; astrocytoma ; post transplant lymphoproliferative disease ; bone marrow purging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Verotoxin (VT) is involved in the etiology of both hemorrhagic colitis and the hemolytic uremic syndrome which are microvasculopathies of the colon and pediatric renal glomerulus respectively. Thus, VT can be considered a vasotoxin. Cell sensitivity in vitro varies according to the receptor glycolipid (globotriaosyl ceramide-Gb3) expression and also to intracellular trafficking of the receptor/toxin complex, such that in highly sensitive cells, the toxin is targeted to the endoplasmic reticulum and nuclear envelope. Such cells include tumor cells which have become drug resistant. Thus Gb3 is upregulated in certain tumors and when such tumor cells become drug resistant, their sensitivity to verotoxin increases. This may be due to a direct role of the MDR1 drug efflux pump in glycolipid biosynthesis. In addition to the tumor tissue, the toxin receptor may also be expressed in the tumor neovasculature suggesting that activated endothelial cells may be verotoxin sensitive. Thus VT may have both a direct and indirect antineoplastic potential. VT has proved highly effective in a xenograft cancer model and the possible therapeutic use of VT is discussed.
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  • 70
    ISSN: 1573-4935
    Keywords: ceramide glycanase ; cancer cells ; glycosphingolipid ; sphingosine ; ceramide ; apoptosis ; PPMP ; PDMP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Ceramide glycanase (CGase) activities have been detected in different human tumor cells (colon, carcinoma Colo-205; neuroblastoma, IMR-32; breast cancer lines, SKBr3 and MCF7). However, the level of enzymatic activity is lower in these cells compared to that present in other mammalian tissues reported before (Basu, M., Kelly, P., Girzadas, M. A., Li, Z., and Basu, S. Methods Enzymol. (in press)). The majority of CGase activity was found in the 100,000g soluble supernatant fraction isolated from all these cell lines and tissues. Using the soluble enzyme, the requirement for optimum CGase activity was found to be consistent with previous observations found for rat and rabbit tissues (Basu, M., Dastgheib, S., Girzadas, M. A., O'Donnell, P. H., Westervelt, C. W., Li, Z., Inokuchi, J. I., and Basu, S. (1998) Acta Pol. Biochim. 42:327). The CGase activities from both Colo-205 and IMR-32 cells are optimum at a protein to detergent ratio of one. All the mammalian CGases, including human cancer cells, show an optimum pH between 5.5 and 5.8 in sodium acetate buffer. The CGase activities from cancer cells are found to be cation-independent; however, mercury, zinc, and copper ions seem to inhibit the enzyme activity substantially in both tumor cells lines. The mercury ion inhibition of CGase activities from all different sources indicates a possible structural homology in the CGase proteins. Radiolabeled substrates, labeled at the sphingosine double bond or at the 3-position of sphingosine without modifying double bond of sphingosine were used in this investigation. Both were active substrates with all enzyme preparations isolated from different cancer cells (apparent Km, 500 μM for nLcOse5[3H-DT]Cer and 350 μM for GgOse4[sph-3-3H]Cer with Colo-205 enzyme). Structural analogues of ceramide and sphingosine (L-PPMP, L-PDMP, alkylamines, and Tamoxifen) inhibited cancer cell CGase activities in vitro.
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  • 71
    ISSN: 1573-7403
    Keywords: apoptosis ; corticotroph adenomas ; heat shock proteins ; p53
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The pathogenesis of corticotroph adenomas is unknown. In a recent study accumulation of p53 protein was detected by immunohistochemistry in a substantial proportion of pituitary corticotroph adenomas, and it has been suggested that it may be causally related to their development. However, other immunohistochemical studies have not confirmed the high incidence of p53 accumulation in this tumor type. Therefore, in the present study, p53 protein accumulation was re-examined in a series of 31 cases of corticotroph adenomas, using different sets of well validated anti-p53 antibodies. Furthermore, in view of the known association of p53 protein with apoptosis, and the known property of p53 to form complexes with heat shock proteins (HSPs), the relationship of p53 accumulation in corticotroph adenomas with apoptosis and HSP-70 was also investigated. Tumor samples staining positively for ACTH from patients with Cushing's disease or Nelson's syndrome were studied. Accumulation of p53 protein was tested by the standard ABC method using two different sets of clone Pab1801 and DO-7 monoclonal antibodies, applied after incubation of sections in a microwave oven. Using the DO-7 antibody, nuclear accumulation of p53 protein was detected in a total of 15 cases, with cytoplasmic staining observed in only 3 tumors. In contrast, using the Pab1801 antibody nuclear staining was observed in only 5 adenomas, with 11 adenomas demonstrating focal cytoplasmic immunoreactivity. Parallel sections of all corticotroph tumors demonstrating cytoplasmic accumulation of p53 protein were tested for the immunohistochemical presence of heat shock protein HSP-70. A striking similar distribution pattern of these two proteins was observed. Apoptosis, identified by the in situ end labeling technique, was detected in a total of 15 out of 28 corticotroph adenomas tested. Calculation of the apoptotic labeling index (ALI) by image analysis showed a significantly lower ALI in those corticotroph adenomas demonstrating nuclear p53 accumulation compared to those with no nuclear p53 immunostaining (p〈0.05). There was no significant difference in the ALI between cytoplasmic p53 positive and negative tumors. It is concluded that depending on the antibody used there is a significant variation of p53 protein detection in corticotroph adenomas. Overall, a significant proportion of corticotroph adenomas studied expressed the p53 protein, which depending on the antibody used, was located either in the nucleus and/or the cytoplasm of tumorous corticotroph cells. Cytoplasmic accumulation of p53, as shown by our colocalization studies with HSP-70, may be due to p53/HSP-70 complex formation. Although such a complex-mediated cytoplasmic exclusion of p53 has no significant effect on apoptosis, nuclear accumulation of p53 protein is associated with a significantly lower apoptotic index indicating a failure of p53 protein to exert its apoptotic action in at least a subset of this tumor type.
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  • 72
    ISSN: 1573-675X
    Keywords: Adenosine ; apoptosis ; necrosis ; physiopathological implications.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Adenosine modulates the proliferation, survival and apoptosis of many different cell types, ranging from epithelial, endothelial and smooth muscle cells, to cells of the immune and neural lineages. In this review, we critically discuss the available in vitro and in vivo data which support a role for adenosine in both development-associated apoptosis, and in diseases characterized by either pathologically increased cell death (e.g., ischemia, trauma and aging-associated neurodegeneration) or abnormally reduced spontaneous apoptosis (e.g., cancer). Particular emphasis is given to the possible role of extracellular adenosine receptors, since these may represent novel and attractive molecular targets for the pharmacological modulation of apoptosis. In some instances, adenosine-induced cell death has been demonstrated to require entry of the nucleoside inside cells; however, in many other cases, activation of specific adenosine extracellular receptors has been demonstrated. Of the four G protein-coupled adenosine receptors so far identified, the A2A and the A3 receptors have been specifically implicated in modulation of cell death. For the A3 receptor, results obtained by exposing both cardiomyocytes and brain astrocytes to graded concentrations of selective agonists suggest induction of both cell protection and cell death. Such opposite effects, which likely depend on the degree of receptor activation, may have important therapeutic implications in the pharmacological modulation of cardiac and brain ischemia. For the A2A receptor, recent intriguing data suggest a specific role in immune cell death and immunosuppression, which may be relevant to both adenosine-deaminase-immunodeficiency syndrome (a pathology characterized by accumulation of adenosine to toxic levels) and in tumors where induction of apoptosis via activation of specific extracellular receptors may be desirable. Finally, preliminary data suggest that, in a similar way to the adenosine-deaminase-immunodeficiency syndrome, the abnormal accumulation of adenosine in degenerative muscular diseases may contribute to muscle cell death. Although the role of adenosine receptors in this effect still remains to be determined, these data suggest that adenosine-induced apoptosis may also represent a novel pathogenic pathway in muscular dystrophies.
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  • 73
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    Apoptosis 4 (1999), S. 317-319 
    ISSN: 1573-675X
    Keywords: Anti-tumor therapy ; Apoptin® ; apoptosis ; Bcl-2 ; p53.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Apoptin, a protein encoded by an avian virus, induces apoptosis in various cultured human tumorigenic and/ or transformed cell lines, e.g. derived from breast and lung tumor, leukemia, lymphoma, osteosarcoma melanoma, cholangiocarcinoma, and hepatoma. In such cells, Apoptin induces p53-independent apoptosis, and the proto-oncogene Bcl-2 can accelerate this effect. The latter is surprising for, in general, Bcl-2 is known to inhibit e.g., p53-induced apoptosis. On the other hand, in normal non-transformed human cells, Apoptin is unable to induce apoptosis, even when Bcl-2 is over-expressed. In animal models Apoptin-induced apoptosis appears to be a safe and efficient anti-tumor agent. These data, in continuation with the observations that Apoptin is specifically stimulated by Bcl-2 in tumor cells, does not need p53, and is not inhibited by Bcr-Abl in these cells, imply that Apoptin is a potential anti-tumor therapy.
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  • 74
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    Apoptosis 4 (1999), S. 229-234 
    ISSN: 1573-675X
    Keywords: Adenovirus ; apoptosis ; Bcl-2 ; p53 ; Rb ; ventricular myocytes.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Apoptosis or programmed cell death is an important physiologic event crucial for the selective removal of damaged or unwanted cells from body tissues. In the cardiovascular system, apoptosis has been observed in the vasculature and myocardium. Untimely or inappropriate myocardial cell loss through an apoptotic process may contribute to ventricular remodeling and the ultimate demise of ventricular function following injury. Therapeutic interventions designed to modulate or prevent myocardial apoptotic cell loss may therefore prove beneficial in maintaining cardiac function. Incite into the molecular mechanisms that govern apoptosis in mammalian cells has led to the identification of several key factors that promote or prevent the apoptotic process. In this report, we discuss putative regulators of cardiac cell apoptosis with specific reference to the tumor suppressor proteins, p53 and Rb. The interplay between these factors, as well as the anti-apoptotic molecules related to the Bcl-2 the family are discussed in the context of the heart under normal and disease conditions.
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  • 75
    ISSN: 1573-675X
    Keywords: Anti-microtubule agents ; apoptosis ; doxorubicin ; neuroblastoma ; tau ; taxoid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Paclitaxel and docetaxel are potent anti-microtubule and antimitotic agents that induce apoptosis in bone marrow-derived cells and epithelial cells. This study examined apoptosis induced by anti-microtubule agents in the neuroblastoma SK-N-SH cell line with a special focus on tau protein which is one of the main Microtubule-Associated- Proteins (MAPs) in neuronal cells. In time, treatment with 1 μM paclitaxel successively induced formation of bundles, then pseudo-asters concomitantly with mitotic block and phosphorylation of bcl-2 (48 h), then phosphorylation of tau and externalization of phosphatidylserine at the early phase of apoptosis (72 h) and finally DNA fragmentation (96 h). Similar results were obtained with 0.5 μM vinorelbine. Paclitaxel induced a lower increase in tau phosphorylation in differentiated SK-N-SH/RA+ cells which are less sensitive to apoptosis. Moreover, doxorubicin whose mechanism of action is independent of microtubules also induced immunostaining of tau at 72 h treatment. In conclusion, our results on neuroblastoma cells show that overexpression of hyperphosphorylated tau is involved in the apoptotic process induced by anti-microtubule agents and may be extended to others cytostatic drugs. Thus, tau protein may play a role in the cellular events observed in neuroblastoma cells undergoing apoptosis.
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  • 76
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    Apoptosis 4 (1999), S. 71-80 
    ISSN: 1573-675X
    Keywords: abl ; apoptosis ; interleukin-3 ; oncogenes ; ras.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Growth factors promote cell survival and proliferation. Homeostasis is maintained by programmed cell death which occurs when the growth stimulus is withdrawn, in response to negative growth regulators such as interferons, TNF-α and CD95 ligand, or following differentiation. Although acutely-transforming oncogenes often overcome the need for growth factors, growth regulatory cytokines can influence proliferative responses of transformed cells. In this study we investigated the effects of IL-3 on the proliferative responses of parental bone marrow-derived 32D cells and cells transformed with ras and abl oncogenes. We show that treatment of ras-transformed 32D cells with IL-3 reduced proliferative responses and decreased colony-forming ability. These effects were exacerbated in the absence of serum and associated with inhibition of tyrosine kinase activity, down-regulation of RAS and MYC expression, and induction of apoptosis as indicated by DNA fragmentation. In contrast, treatment of parental 32D cells with IL-3, which is obligatory for cell survival and proliferation, increased tyrosine kinase activity, upregulated MYC and RAS expression and maintained DNA integrity. With abl-transformed cells, proliferation and colony-forming ability were also inhibited by IL-3. Tyrosine kinase activity and MYC expression were reduced, but early apoptosis was not evident. Calcium uptake however, was stimulated by IL-3 in both parental and oncogene-transformed cells. These results suggest that threshold levels of tyrosine kinase activity are necessary for cell survival and proliferation and that with ras-transformed cells, IL-3 treatment may result in this threshold being breached. We conclude that in some situations, growth-promoting cytokines can inhibit proliferation of transformed cells and induce cell death by apoptosis.
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  • 77
    ISSN: 1573-675X
    Keywords: ameloblasts ; amelogenesis ; apoptosis ; insulin-like growth factor.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Enamel-producing cells (ameloblasts) pass through several phenotypic and functional stages during enamel formation. In the transition between secretory and maturation stages, about one quarter of the ameloblasts suddenly undergo apoptosis. We have studied this phenomenon using the continuously erupting rat incisor model. A special feature of this model is that all stages of ameloblast differentiation are presented within a single longitudinal section of the developing tooth. This permits investigation of the temporal sequence of gene and growth factor receptor expression during ameloblast differentiation and apoptosis. We describe the light and electron microscopic morphology of ameloblast apoptosis and the pattern of insulin-like growth factor-1 receptor expression by ameloblasts in the continuously erupting rat incisor model. In the developing rat incisor, ameloblast apoptosis is associated with downregulated expression of the insulin-like growth factor-1 receptor. These data are consistent with the hypothesis that ameloblasts are “hard wired” for apoptosis and that insulin-like growth factor-1 receptor expression is required to block the default apoptotic pathway. Possible mechanisms of insulin-like growth factor-1 inhibition of ameloblast apoptosis are presented. The rat incisor model may be useful in studies of physiological apoptosis as it presents apoptosis in a predictable pattern in adult tissues.
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  • 78
    ISSN: 1573-675X
    Keywords: AKR lymphoma malignancy variants ; apoptosis ; apoptosis-related gene expression ; tumor progression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Resistance to apoptosis may be related to tumor progression, due to the implications it might have on both tumor mass and genetic instability. We compared the tendency to spontaneous apoptosis and the proliferative capacity of metastatic growths of several AKR lymphoma variants (TAU-45, TAU-47, TAU-44, TAU-33, TAU-42 and TAU-46, in the order of increasing metastatic potential). We further compared the expression of several apoptosis-related genes. Cell proliferative capacity did not appear to determine malignant behavior since, on the whole, a decrease in S + G2M fraction was observed with increasing malignancy. Sensitivity to apoptotic cell death decreased with increasing malignancy when comparing the TAU-45, TAU-47, TAU-44 and TAU-33 variants, suggesting a role of reduced apoptosis in this T-cell lymphoma. An increase in Bcl-2 content with increasing aggressiveness among these variants, implicates this protein in this tumor progression-related resistance to apoptosis. However, the two variants of highest malignancy, TAU-42 and TAU-46, did not follow the same trend, since they displayed a relatively high content in apoptotic cells and a low Bcl-2 content. Fas receptor expression did not correlate with tendency to apoptosis, indicating that malignant behavior in the AKR lymphoma does not depend on CD95/Fas/APO1 downregulation. Overexpression of p53 was observed only in one of the variants of lowest malignancy.
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  • 79
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    Apoptosis 4 (1999), S. 419-427 
    ISSN: 1573-675X
    Keywords: Anti-oxidant defence ; apoptosis ; ether lipids ; glutathione ; ionising radiation ; stress.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have investigated the mechanisms of induction of apoptosis by the antineoplastic ether lipid ET-18-OCH3 (ALP) in sensitive S49wt mouse lymphoma cells and ALP-resistant S49ar variants, both with wild-type p53, and in related L1210 cells with mutated p53. Ether lipid-resistant S49ar cells were cross-resistant to extracellular stress factors (cold shock, heat shock, H2O2, dimethylsulfoxide) and to radiation-induced apoptosis but not to physiological apoptotic signals (dexamethasone, growth factor deprivation, thapsigargin, C2-ceramide) and expressed similar levels of the apoptosis-regulating proteins Bcl-2, Bcl-X, Bax, Bad and Bak as did the parent S49wt cells. The uptake of [3H]-ALP was strongly reduced in the stress-resistant cells but this was not associated with significant differences in membrane cholesterol:phospholipid content nor in membrane microviscosity. In S49ar cells the activity of the antioxidant enzyme glutathione peroxidase (GSH-Px) was increased 4-fold and depletion of glutathione with the drug L-buthionine-S-R-sulfoximine (L-BSO) lowered the resistance of S49ar cells to ALP, stress factors and ionising radiation. The results indicate that ether lipids induce apoptosis by imposing a special form of physico-chemical stress, mediated by reactive oxygen species but independent of p53 status. The capacity of glutathione-dependent anti-oxidant defence appeared an important and shared determinant of the sensitivity to ether lipids, several types of extracellular stress and ionising radiation.
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  • 80
    ISSN: 1573-675X
    Keywords: Annexin V ; apoptosis ; flow cytometry ; intracellular Ca2+ ; intracellular pH ; mitochondrial membrane potential.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The sequential occurrence of plasma and mitochondrial membrane alterations, intra-cellular pH shifts and changes in intracellular Ca2+ concentration after induction of cell death was monitored by flow cytometry in Jurkat and HSB2-cells. Cell death was induced by treatment with anti-Fas antibodies or by irradiation. Phosphatidylserine (PS) exposure and plasma membrane integrity were measured with FITC-Annexin V adhesion and by Propidium Iodide exclusion. Transition of the mitochondrial membrane potential was monitored by the occurrence of decay of DiOC6 fluorescence. Intracellular pH shifts were monitored by changes in the ratio of fluorescence at 575 nm and at 635 nm of SNARF-1-AM. Fluctuations in intracellular Ca2+ concentration were established by changes in Fura red quenching. The Jurkat cells were sensitive to anti-Fas treatment, while HSB-2 cells were not. HSB-2 cells appeared more sensitive to radiation damage than Jurkat cells. In all experiments the transition of mitochondrial membrane potential occurred first, almost immediately followed by PS exposure. Fluctuations in intracellular Ca2+ concentration occurred later and were less outspoken. A decrease in intracellular pH occurred not earlier than 24 hours after anti-Fas treatment. Chelation of intracellular Ca2+ concentration with BAPTA-AM had no effect on the time sequence of cell death related events.
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  • 81
    ISSN: 1573-904X
    Keywords: MAPK ; caspases ; chemopreventive agents ; phase II drug metabolizing enzymes ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract In therapeutic response to drugs, the plasma concentration range leads to the establishment of a safe and effective dosage regimen. Our hypothesis is that by studying drug concentration-dependent effect on signal transduction mechanisms, a better understanding of the beneficial pharmacodynamic and adverse toxicodynamic responses elicited by the drug may be achieved. Using two classes of chemopreventive compounds (phenolic antioxidants and isothiocyanates), we illustrate the potential utility of two signal transduction pathways elicited by these agents to predict the pharmacodynamic effect (induction of Phase II drug metabolizing enzymes) and the potential toxicodynamic response (stimulation of caspase activity and cytotoxic cell death). At lower concentration, phenolic antioxidants and isothiocyanates activate mitogen-activated protein kinase (MAPK; extracellular signal-regulated protein kinase 2, ERK2; and c-Jun N-terminal kinase 1, JNK1) in a concentration-and time-dependent manner. The activation of MAPK by these compounds may lead to the induction of cell survival/protection genes such as c-jun, c-fos, or Phase II drug metabolizing enzymes. However, at higher concentrations, these agents activate another signaling molecule, ICE/Ced3 cysteine protease enzymes (caspases) leading to apoptotic cell death. The activation of these pathways may dictate the fate of the cells/tissues upon exposure to drugs or chemicals. At lower concentrations, these compounds activate MAPK leading to the induction of Phase II genes, which may protect the cells/tissues against toxic insults and therefore may enhance cell survival. On the other hand, at higher concentrations, these agents may activate the caspases, which may lead to apoptotic cell death, and have toxicity. Understanding the activation of these and other signal transduction events elicited by various drugs and chemicals may yield insights into the regulation of gene expression of drug metabolizing enzymes and cytotoxicity. Thus, the study of signaling events in cell survival (hemeostasis) and cell death (cytotoxicity) may have practical application during pharmaceutical drug development.
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  • 82
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    Plant growth regulation 27 (1999), S. 119-123 
    ISSN: 1573-5087
    Keywords: apoptosis ; carrot protoplast ; DNA ladder ; ethrel ; ethylene ; nucleus condensation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In recent years, apoptosis has been reported to exist in plants during normal development and in response to stress. However, little is known about the relation of hormones to this form of programmed cell death. Here, we report examination of characteristics of apoptosis in carrot protoplasts induced by ethylene evolved from ethrel (2-chloroethylphosphonic acid). Nucleus condensation and DNA ladders were observed, and neutral comet assay, which detects DNA cleavage, also provided evidence that ethrel treatment resulted in nuclear DNA fragmentation. Strikingly, a close correlation between the incidence of DNA comets and the percentage of apoptotic protoplasts was shown in ethrel-treated carrot protoplasts. In conclusion, this study demonstrated that ethylene is an active inducer of apoptosis in carrot protoplasts, and that ethylene-induced plant cell death showed characteristics similar to those of apoptosis in animals.
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  • 83
    ISSN: 1573-6903
    Keywords: Ethanol ; apoptosis ; gangliosides ; LIGA20 ; cerebellar granule cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Ethanol significantly enhances cell death of differentiated rat cerebellar granule neurons on culture in a serum-free medium containing a depolarizing concentration of KCl (25 mM), 5 μM MK-801 (an NMDA receptor antagonist), and 20–200 mM ethanol for 1–4 days. Cell death augmented by ethanol was concentration- and time-dependent with neurons displaying hallmark apoptotic morphology and DNA fragmentation that correlated with the activation of cytosolic caspase-3. Inclusion of 5 μM MK-801 or 100 μM glycine in culture media did not alter rates of cell death indicating ethanol toxicity is mediated via an NMDA receptor-independent pathway. Preincubation with 50 μM gangliosides GM1, GD1a, GD1b or GT1b for 2 h, or preincubation with 10 μM LIGA20 (a semisynthetic GM1 with N-dichloroacetylsphingosine) for 10 min, attenuated caspase-3 activity and ethanol-induced cell death. Data show native gangliosides and a synthetic derivative are potently neuroprotective in this model of ethanol toxicity, and potentially serve as useful probes to further unravel the mechanisms relevant to neuronal apoptosis.
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  • 84
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    Journal of bioenergetics and biomembranes 31 (1999), S. 291-304 
    ISSN: 1573-6881
    Keywords: Cell death ; aging ; necrosis ; apoptosis ; mitochondria ; oxidative phosphorylation ; electron transport chain ; ATP synthase ; cytochrome c ; mitochondrial DNA ; reactive oxygen species (ROS)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Traditionally, mitochondria have been viewed as the “powerhouse” of the cell, i.e., the site of theoxidative phosphorylation machinery involved in ATP production. Consequently, much of theresearch conducted on mitochondria over the past 4 decades has focused on elucidating both thosemolecular events involved in ATP synthesis by oxidative phosphorylation and those involved inthe biogenesis of the oxidative phosphorylation machinery. While monumental achievements havebeen made, and continue to be made, in the study of these remarkable but extremely complexprocesses essential for the life of most animal cells, it has been only in recent years that a largebody of biological and biomedical scientists have come to recognize that mitochondria participatein other important processes. Two of these are cell death and aging which, not surprisingly, are relatedprocesses both involving, in part, the oxidative phosphorylation machinery. This new awareness hassparked a new and growing area of mitochondrial research, that has become of great interest to awide variety of scientists ranging from those involved in elucidating the role of mitochondria incell death and aging to those interested in either suppressing or facilitating these processes as itrelates to identifying new therapies or drugs for human disease. It is the purpose of this briefintroductory review to provide an overview of those mitochondrial events involved in the life anddeath of animal cells and to indicate how these events might relate to the human aging process.Much more is known, much remains controversial, and even more remains to be learned as indicatedin the excellent set of minireviews that follow.
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  • 85
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    Journal of bioenergetics and biomembranes 31 (1999), S. 321-326 
    ISSN: 1573-6881
    Keywords: Mitochondria ; apoptosis ; caspases ; cytochrome c ; Fas ; bcl-2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract In the past few years, it has become widely appreciated that apoptotic cell death generallyinvolves activation of a family of proteases, the caspases, which undermine the integrity ofthe cell by cleavage of critical intracellular substrates. Caspases, which are synthesized asinactive zymogens, are themselves caspase substrates and this cleavage leads to their activation.Hence, the potential exists for cascades of caspases leading to cell death. However, it has beenrecently recognized that another, perhaps more prominent route to caspase activation, involvesthe mitochondria. Upon receipt of apoptotic stimuli, either externally or internally generated,cells initiate signaling pathways which converge upon the mitochondria to promote release ofcytochrome C to the cytoplasm; cytochrome c, thus released, acts as a potent cofactor incaspase activation. Even cell surface “death receptors” such as Fas, which can trigger directcaspase activation (and potentially a caspase cascade), appear to utilize mitochondria as partof an amplification mechanism; it has been recently demonstrated that activated caspases cancleave key substrates to trigger mitochondrial release of cytochrome c, thereby inducing furthercaspase activation and amplifying the apoptotic signal. Therefore, mitochondria play a centralrole in apoptotic cell death, serving as a repository for cytochrome c.
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  • 86
    ISSN: 1573-6881
    Keywords: Energy expenditure ; reactive oxygen species ; cellular viability ; apoptosis ; necrosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Uncoupling proteins, a subgroup of the mitochondrial anion transporter superfamily, have beenidentified in prokaryotes, plants, and mammalian cells. Evolutionary conservation of thesemolecules reflects their importance as regulators of two critical mitochondrial functions, i.e.,ATP synthesis and the production of reactive oxygen species (ROS). Although the amino acidsequences of the three mammalian uncoupling proteins, UCP1, UCP2 and UCP3, are verysimilar, each homolog is the product of a unique gene and important differences have beendemonstrated in their tissue-specific expression and regulation. UCP1 and UCP3 appear to bekey regulators of energy expenditure, and hence, nonshivering thermogenesis, either in brownadipose tissue (UCP1) or skeletal muscle (UCP3). UCP2 is expressed more ubiquitously,although generally at low levels, in many tissues. There is conflicting evidence about itsimportance as a regulator of resting metabolic rate. However, evidence suggests that thishomolog might modulate the mitochondrial generation of ROS in some cell types, includingmacrophages and hepatocytes. While the induction of various uncoupling protein homologsprovides adaptive advantages, both to the organism (e.g., thermogenesis) and to individual cells(e.g., reduced ROS), increased uncoupling protein activity also increases cellular vulnerability tonecrosis by compromising the mitochondrial membrane potential. This narrow “risk—benefit”margin necessitates tight control of uncoupling protein activity in order to preserve cellularviability and much remains to be learned about the regulatory mechanisms involved.
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  • 87
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    Cell biology and toxicology 15 (1999), S. 367-373 
    ISSN: 1573-6822
    Keywords: apoptosis ; drugs ; hepatitis ; mitochondria ; steatosis ; steatohepatitis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Mitochondria are involved in fatty acid β-oxidation, the tricarboxylic acid cycle, and oxidative phosphorylation, which provide most of the cell energy. Mitochondria are also the main source of reactive oxygen species in the cell and are involved in cell demise through opening of the mitochondrial permeability transition pore. It was therefore to be expected that mitochondrial dysfunction could be a major mechanism of drug-induced liver disease. Microvesicular steatosis (which may cause liver failure, coma, and death) is the consequence of severe impairment of mitochondrial β-oxidation. Endogenous compounds (such as cytokines or female sex hormones) or xenobiotics (including toxins such as ethanol and drugs such as aspirin, valproic acid, ibuprofen, or zidovudine) can inhibit β-oxidation directly or through a primary effect on the mitochondrial genome or the respiratory chain itself. In some patients, infections and cytokines, or inborn errors of β-oxidation enzymes or the mitochondrial genome, may favor the appearance of drug-induced microvesicular steatosis. Nonalcoholic steatohepatitis may develop under conditions causing prolonged, microvesicular, and/or macrovacuolar steatosis. In this condition, chronic impairment of mitochondrial β-oxidation (causing steatosis) and the respiratory chain (increasing the production of ROS) lead to lipid peroxidation, which, in turn, may cause the diverse lesions of steatohepatitis, namely, necrosis, inflammation, Mallory's bodies, and fibrosis. Finally, mitochondria are involved in several forms of drug-induced cytolytic hepatitis, through inhibition or uncoupling of respiration or through a drug-induced or reactive metabolite-induced mitochondrial permeability transition. The latter effect commits hepatocytes to either apoptosis or necrosis, depending on the number of organelles that have undergone the permeability transition.
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  • 88
    ISSN: 1573-6830
    Keywords: Parkinson's disease ; catecholamines ; oxidative metabolites ; phosphorylation ; DNA damage ; apoptosis ; p53
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 1. The pathogenesis of the selective degeneration of the dopaminergic neurons in Parkinson's disease is still enigmatic. Recently we have shown that dopamine can induce apoptosis in postmitotic neuronal cells, as well as in other cellular systems, thus suggesting a role for this endogenous neurotransmitter and associated oxidative stress in the neuronal death process. 2. Dopamine has been shown to be capable of inducing DNA damage through its oxidative metabolites. p53 is a transcription factor that has a major role in determining cell fate in response to DNA damage. We therefore examined the possible correlation between dopamine-triggered apoptosis, DNA damage and levels of total phosphorylated p53 protein in cultured mouse cerebellar granule neurons. 3. Marked DNA damage and apoptotic nuclear condensation and fragmentation were detected within several hours of exposure to dopamine. An associated marked threefold increase in p53 phosphorylation was observed within this time window. Using a temperature-sensitive p53 activation system in leukemia LTR6 cells, were found that p53 inactivation dramatically attenuated dopamine toxicity. 4. We therefore conclude that DNA damage and p53 activation may have a role in mediating dopamine-induced apoptosis. Modulation of the p53 system may therefore have a protective role against the toxicity of this endogenous neurotransmitter and associated oxidative stress.
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  • 89
    ISSN: 1573-7217
    Keywords: apoptosis ; Bax ; Bcl‐2 ; breast cancer ; chemotherapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Optimizing chemotherapeutic drug delivery strategies relies, in part, on identification of the most clinically effective sequence, dose, and duration of drug exposure. The combination of dose intensive etoposide (VP‐16) followed by cyclophosphamide has clinical efficacy in the treatment of advanced breast cancer. However, molecular mechanisms that underlie the effectiveness of this combination of chemotherapeutic agents have not been investigated. In this study we investigated regulation of BAX and BCL‐2 expression by VP‐16 and cyclophosphamide as a potential mechanism for the induction of breast cancer cell death induced by this regimen. There was a dose and time dependent increase in BAX expression in the breast cancer cell lines MCF‐7, MDA‐MB‐435S, and MDA‐MB‐A231 following in vitro treatment with 50–100 μM VP‐16. Elevation of BAX protein expression in the presence of VP‐16 alone did not correlate with reduced viability or induction of apoptosis in MCF‐7, MDA‐MB‐435S, or MDA‐MB‐A231. VP‐16 did effectively block the breast cancer cell lines evaluated (MCF‐7 and MDA‐MB‐435S) at G2/M phase of the cell cycle, confirming activity of the drug in vitro. MCF‐7 and MDA‐MB‐435S cells that were pre‐treated with VP‐16 and subsequently exposed to 1.0–12.0 μg/m1 4‐hydroperoxycyclophosphamide (4HC), an active metabolite of cyclophosphamide, had markedly reduced viability when compared to matched controls treated with either VP‐16 or 4HC individually. Consistent with this loss of viability, exposure of all three cell lines to the combination of VP‐16 and 4HC resulted in higher BAX protein levels than those observed following treatment with either single agent. This combination of chemotherapeutic agents also resulted in reduced BCL‐2 expression. These observations suggest that combination chemotherapy may derive its efficacy, in part, through coordinated regulation of specific gene products associated with apoptosis. Characterization of molecular events that underlie susceptibility of specific tumor cells to combination chemotherapeutic regimens may lead to additional improvements in treatment strategies for this disease.
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  • 90
    ISSN: 1573-7217
    Keywords: apoptosis ; breast cancer ; doxorubicin ; hsp27 ; topoisomerase II
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Previously we demonstrated that heat shock protein 27 (hsp27) overexpression confers resistance to the chemotherapeutic agent doxorubicin in MDA–MB–231 breast cancer cells. Since induction of apoptosis is one underlying mechanism of chemotherapeutic drug action, we investigated the effect of hsp27 overexpression on doxorubicin–induced apoptosis, finding that hsp27 protects MDA–MB–231 cells from apoptosis. We also examined expression of the doxorubicin target, topoisomerase II (topo II), in control and hsp27–overexpressing stable transfectants, as topo II expression is important for both drug sensitivity and the initiation of apoptosis by doxorubicin. The relative levels of both topo IIα and β were higher in the controls than the hsp27–overexpressing clones, suggesting that the apoptotic protective effect of hsp27 overexpression in MDA–MB–231 cells is associated with altered topo II expression.abstract
    Type of Medium: Electronic Resource
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  • 91
    ISSN: 1573-7217
    Keywords: angiogenesis inhibitor ; apoptosis ; Bcl‐2 ; breast cancer ; eicosapentaenoic acid ; TNP‐470
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effects of eicosapentaenoic acid (EPA) and an angiogenesis inhibitor (TNP‐470) on the suppression of breast cancer cell growth were examined in five human breast cancer cell lines (MDA‐MB‐231, T‐47D, MCF‐7, KPL‐1, and MKL‐F). In all five cell lines, EPA and TNP‐470 alone both showed tumor growth inhibition in a time‐ and dose‐dependent manner, and in combination, a synergistic effect was seen at high concentrations. EPA plus TNP‐470 treatment evoked apoptosis as confirmed by the appearance of sub G1 populations, by DNA fragmentation, and by cell morphology. With the combination, the expression of Bax and Bc1‐xS, the apoptosis‐enhancing proteins, was more up‐regulated and that of Bcl‐2 and Bcl‐xL, the apoptosis‐suppressing proteins, was more down‐regulated compared to the use of EPA or TNP‐470 alone, suggesting that their synergistic effect was due to an acceleration of apoptosis.
    Type of Medium: Electronic Resource
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  • 92
    ISSN: 1573-7217
    Keywords: apoptosis ; glycogen‐rich breast carcinoma ; prognosis ; proliferation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We determined the proliferation rate and apoptotic activity of glycogen‐rich carcinomas of the breast as opposed to non‐clear cell tumors by means of MIB‐1 immunohistochemistry and in situ detection of oligonucleosomal fragments (TUNEL reaction). The retrospective biopsy series included six invasive clear cell carcinomas of the glycogen‐rich type as well as 15 randomly selected cases of invasive ductal carcinoma without evidence of glycogen storage. Three patients in the clear cell group and seven patients in the control cohort developed lymph‐node metastasis. The MIB‐1 labeling index of glycogen‐rich carcinomas averaged 9.05%, while that of the controls was 30.03%. Apoptotic nuclei were present in a mean of 1.26% of glycogen‐rich carcinoma cells. The control tumors exhibited an average apoptotic frequency of 5.85%. Tumor size, hormone receptor status, and presence or absence of lymph node involvement were found not to correlate with either proliferation or apoptosis. We conclude that glycogen‐rich breast carcinomas are characterized by a peculiar ‘low proliferation‐low apoptosis’ cell kinetic profile. The aggressive clinical behavior of these neoplasms may possibly be accounted for by an ineffective apoptotic elimination of otherwise slowly proliferating tumor cells.
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  • 93
    ISSN: 1573-6903
    Keywords: Calpain ; caspases ; maitotoxin ; necrosis ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Maitotoxin is a potent toxin that activates voltage and receptor-mediated Ca2+ channels, resulting in Ca2+ overload and rapid cell death. We report that maitotoxin-induced cell death is associated with activation of calpain but not caspase-3 proteases in septo-hippocampal cell cultures. Calpain and caspase-3 activation were examined by accumulation of protease-specific breakdown products to α-spectrin. Cell death manifested exclusively necrotic-like characteristics including round, shrunken nuclei, even distribution of chromatin, absence of DNA fragmentation and failure of protein synthesis inhibition to reduce cell death. Necrotic cell death was observed in neurons and astroglia. Calpain inhibitor II inhibited calpain-specific processing of α-spectrin and significantly reduced cell death. The pan-caspase inhibitor, Z-D-DCB, nominally attenuated cell death. Results suggest that: (1) calpain, but not caspase-3, is activated as a result of maitotoxin-induced Ca2+ influx; (2) necrotic cell death caused by maitotoxin exposure is partially mediated by calpain activation; (3) maitotoxin is a useful tool to investigate pathological mechanisms of necrosis.
    Type of Medium: Electronic Resource
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  • 94
    ISSN: 1573-7217
    Keywords: apoptosis ; Bcl-2 ; breast cancer ; c-Myc ; metastasis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The overexpression of Bcl-2, an anti-apoptotic oncogene, identifies human T1 breast cancer patients who have an increased risk of lymph-node metastasis. We examined in these patients (n=142) whether the c-Myc oncogene influences metastatic progression in conjunction or not with Bcl-2 expression and the loss of apoptosis in tumors. The association between Bcl-2 and lymph-node metastasis was only significant when c-Myc was concomitantly expressed (χ2 test, p=0.008). Moreover, very large associations (pOR=6.4) between c-Myc and lymph-node metastasis were observed among Bcl-2 positive tumors and tumors with loss of apoptosis (pOR=8.4). In contrast, the metastatic advantage linked to Bcl-2 was decreased (pOR=2) when c-Myc was not coexpressed. It is concluded that the synergism between Bcl-2 and c-Myc oncogenes may promote metastasis in breast tumors, linked to loss of apoptosis.
    Type of Medium: Electronic Resource
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  • 95
    ISSN: 1573-7217
    Keywords: apoptosis ; breast cancer ; doxorubicin ; IGF‐I ; paclitaxel
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Insulin‐like growth factor (IGF)‐I protects many cell types from apoptosis. As a result, it is possible that IGF‐I‐responsive cancer cells may be resistant to apoptosis‐inducing chemotherapies. Therefore, we examined the effects of IGF‐I on paclitaxel and doxorubicin‐induced apoptosis in the IGF‐I‐responsive breast cancer cell line MCF‐7. Both drugs caused DNA laddering in a dose‐dependent fashion, and IGF‐I reduced the formation of ladders. We next examined the effects of IGF‐I and estradiol on cell survival following drug treatment in monolayer culture. IGF‐I, but not estradiol, increased survival of MCF‐7 cells in the presence of either drug. Cell cycle progression and counting of trypan‐blue stained cells showed that IGF‐I was inducing proliferation in paclitaxel‐treated but not doxorubicin‐treated cells. However, IGF‐I decreased the fraction of apoptotic cells in doxorubicin‐ but not paclitaxel‐treated cells. Recent work has shown that mitogen‐activated protein kinase (MAPK) and phosphotidylinositol‐3 (PI‐3) kinase are activated by IGF‐I in these cells. PI‐3 kinase activation has been linked to anti‐apoptotic functions while MAPK activation is associated with proliferation. We found that IGF‐I rescue of doxorubicin‐induced apoptosis required PI‐3 kinase but not MAPK function, suggesting that IGF‐I inhibited apoptosis. In contrast, IGF‐I rescue of paclitaxel‐induced apoptosis required both PI‐3 kinase and MAPK, suggesting that IGF‐I‐mediated protection was due to enhancement of proliferation. Therefore, IGF‐I attenuated the response of breast cancer cells to doxorubicin and paclitaxel by at least two mechanisms: induction of proliferation and inhibition of apoptosis. Thus, inhibition of IGF‐I action could be a useful adjuvant to cytotoxic chemotherapy in breast cancer.
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  • 96
    Electronic Resource
    Electronic Resource
    Springer
    Angiogenesis 3 (1999), S. 101-116 
    ISSN: 1573-7209
    Keywords: angiogenesis ; apoptosis ; cell death ; endothelial cell ; neovascularization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Angiogenesis, the process of new microvessel development, is encountered in a select number of physiological processes and is central to the pathogenesis of a wide variety of diseases. There is now convincing evidence that regulated patterns of endothelial cell survival and death, a process known as apoptosis, play a central role in the periodic remodeling of the vasculature, and in the timely evolution and regression of angiogenic responses. In this review we discuss the current evidence suggesting a role for inducers and inhibitors of angiogenesis as well as other mediators that modify endothelial cells functions in the survival and death of endothelial cells. We also discuss how dysregulation of apoptosis can lead to aberrant angiogenesis as demonstrated in the pathogenesis of retinopathy of prematurity and cancer.
    Type of Medium: Electronic Resource
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  • 97
    Electronic Resource
    Electronic Resource
    Springer
    Breast cancer research and treatment 54 (1999), S. 269-278 
    ISSN: 1573-7217
    Keywords: apoptosis ; breast carcinoma ; cell cycle ; curcumin ; cytotoxicity ; gene expression ; RT‐PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Curcumin has anti‐inflamatory, antiproliferative, and antitumor effects. To understand the chemopreventive mechanism of curcumin against human malignancies, the cellular and molecular changes induced by this agent in human mammary epithelial (MCF‐10A) and breast carcinoma (MCF‐ 7/TH) cell lines were investigated. The human multidrug‐ resistant breast cancer cell line was 3.5 fold more sensitive to curcumin than the mammary epithelial cell line. Even though both cell lines accumulated a similar amount of curcumin, a significantly higher percentage of apoptotic cells was induced in breast cancer cells compared to a very low percentage of apoptosis in mammary epithelial cells. Incubation of breast cancer cells with 20 and 40 μM curcumin for 24 h induced G2 block and sub‐ G0/G1 cell population, respectively. Curcumin treatment caused a reduction in the expression of Ki67, PCNA, and p53 mRNAs in breast cancer cells. The human mammary epithelial cell line showed a down‐regulation of p21 mRNA and an up‐regulation of Bax mRNA expression with curcumin treatment. The results suggest that apoptosis is involved in the curcumin‐induced inhibition of tumor cell growth, and genes associated with cell proliferation and apoptosis may be playing a role in the chemopreventive action of curcumin. Abbreviations: EGF: epidermal growth factor; D-MEM: Dulbecco' Modified Eagle Medium; EDTA: ethylene diamine tetra‐acetic acid; PBS: phosphate buffered saline; TdT: terminal deoxynucleotidyl transferase; FBS: fetal bovine serum; RT‐PCR: reverse transcription‐polymerase chain reaction; PCNA: proliferating cell nuclear antigen; TNF: tumor necrosis factor; TPA: 12‐tetradecanoyl‐phorbol‐13‐acetate.
    Type of Medium: Electronic Resource
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  • 98
    Electronic Resource
    Electronic Resource
    Springer
    Breast cancer research and treatment 53 (1999), S. 263-269 
    ISSN: 1573-7217
    Keywords: apoptosis ; breast cancer ; neo-adjuvant chemotherapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The use of neo-adjuvant chemotherapy (often referred to as pre-operative or primary chemotherapy) represents a major change in the management of breast cancer as a systemic disease. Laboratory studies have shown that many anti-cancer agents with differing modes of action achieve cytotoxic effects by inducing apoptosis. In this study, we investigated the induction of apoptosis by neo-adjuvant chemotherapy in human breast cancer. The aim was to determine whether a correlation existed between post chemotherapy apoptotic index (AI) and clinical response and patients' survival. Our results indicate that apoptosis is induced by neo-adjuvant chemotherapy and that the response is variable. Our data show that post chemotherapy AI correlated with clinical response and increased patient survival, including both relapse (disease) free survival and overall survival. Post-neo-adjuvant chemotherapy AI levels in primary breast cancer may possibly predict an individual patient's overall response.
    Type of Medium: Electronic Resource
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  • 99
    ISSN: 1573-7217
    Keywords: apoptosis ; breast carcinoma ; immunology ; tolerance/suppression ; tumor immunology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Breast tumors are frequently associated with a predominantly lymphocytic infiltrate, which constitutes an immune response against the tumor. In spite of this massive infiltrate, the immune response appears to be inefficient and the tumor is able to evade it. We propose that in breast cancer, tumor escape from immunological surveillance results from the induction of apoptosis of Fas‐bearing activated lymphocytes by FasL‐bearing breast cancer cells. To test this proposal we studied the expression of FasL by human breast carcinomas and the MCF‐7 breast cancer cell line by RT‐PCR, immunohistochemistry, and Western Blot. Moreover, we describe the presence of apoptosis and Fas expression in the lymphocytic population surrounding the tumor. Strong membranous and cytoplasmic staining was detected in ductal carcinomas and hyperplastic breast tissue, but it was absent from normal breast tissue. No staining was found in normal glands in the non‐tumor quadrant; however, the normal appearing ducts surrounding the carcinoma (tumor quadrant) showed intense immunoreactivity. Apoptosis was found predominantly among the lymphocytic population, as well as in the blood vessels and fibro‐fatty tissue close to the tumor. Further characterization of apoptotic cells demonstrated that they were CD3+ cells. Our results suggest the breast tumors may elude immunological surveillance by inducing, via the Fas‐FasL system, the apoptosis of activated lymphocytes. Recent data have demonstrated FasL RNA in other tumor types. Upregulation of FasL expression in hyperplastic and normal breast ducts close to the tumor also suggests a possible role in early neoplastic transformation and proliferation. Abbreviations: Con A: concanavalin A; FasL: Fas ligand; RT-PCR: reverse transcription+polymerase chain reaction.
    Type of Medium: Electronic Resource
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  • 100
    Electronic Resource
    Electronic Resource
    Springer
    Breast cancer research and treatment 55 (1999), S. 73-83 
    ISSN: 1573-7217
    Keywords: apoptosis ; Bax ; Bcl‐2 ; breast ; chemotherapy ; estradiol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The present study explored the effects of three commonly used chemotherapeutic agents on the Bcl‐2/Bax apoptosis pathway and the interaction of these chemotherapeutic drugs with the estradiol‐mediated regulation of this pathway. Our results showed that: (1) Treatment of MCF‐7 cells with Adriamycin resulted in time‐ and concentration‐dependent decreases in Bcl‐2 and increases in Bax mRNA and protein levels. (2) Camptothecin elicited similar trends on Bcl‐2 and Bax as Adriamycin, while etoposide, at 50–100 fold (1–5 μM) the effective concentration of Adriamycin and camptothecin, only resulted in an increase in Bax mRNA levels. (3) Adriamycin and camptothecin, but not etoposide, were effective in suppressing estradiol‐stimulated increases in Bcl‐2 mRNA levels. Our study provides evidence that the Bcl‐2/Bax apoptosis pathway may be differentially regulated by chemotherapeutic agents. In addition, interaction between these agents and estradiol on the Bcl‐2/Bax apoptosis pathway may also exist.
    Type of Medium: Electronic Resource
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