There is general agreement that apoptosis (programmed/physiological cell death) plays an important role in regulating normal hematopoiesis and that so-called naked nuclei (NN) present end stages of megakaryopoiesis. Using in situ end-labeling techniques (ISEL), a morphometric analysis was performed on bone marrow trephine biopsies repeatedly derived from patients with idiopathic (primary) osteo-/myelofibrosis (IMF). The purpose of this study was to quantify apoptosis and to elucidate its relationship with NN of megakaryopoiesis. Moreover, evolution of this phenomenon during progression of myelofibrosis (assessed by morphometry of silver-stained sequential biopsies) and its predictive value were investigated. In IMF, morphometric measurements revealed a rate of ISEL-positive cells that was significantly higher than previously published data on the normal bone marrow and in patients with acute myeloblastic leukemia, but lower than the myelodysplastic syndromes. However, NN with their tightly packed (nuclear) lobules and condensed chromatin were not reactive. Statistical evaluation failed to show a significant correlation between incidence of apoptotic cells and fiber density or an increase in this feature associated with progression of fibro-osteosclerotic marrow changes and prognosis. The failure of NN to react in ISEL implicates an absence of DNA fragmentation characteristic of apoptosis. For this reason our findings are in keeping with the assumption that NN are compatible with para-apoptosis.

A comparative morphometric analysis was performed on smears and trephine biopsies of normal bone marrow and in chronic myelogenous leukaemia (CML) to assess the effects of therapy on apoptosis and cell proliferation. The in situ end‐labelling (ISEL) technique was used for the demonstration of programmed cell death, in combination with the monoclonal antibody PG‐M1 to identify macrophages. Cell proliferation was evaluated by employing the monoclonal antibody PC10 directed against proliferating cell nuclear antigen (PCNA). In CML (48 patients), significantly higher rates of apoptosis were observed than in normal bone marrow (smears, frozen sections, and paraffin‐embedded samples) of 15 patients. In contrast, the PCNA labelling index of CML was not different from controls. In bone marrow tissue derived from CML patients, about 36 per cent of apoptotic bodies were ingested with CD68‐positive macrophages. Study of the histotopographical distribution of labelled cells revealed that in CML, in contrast to the normal bone marrow, programmed cell death and PCNA activity were concentrated along the paratrabecular generation zone. In 28 patients with CML treated with interferon (IFN), sequential trephine biopsies displayed a significant enhancement of apoptosis which was associated with a decrease in PCNA reactivity. In contrast to this finding, no such alterations could be observed in 24 patients who received busulfan (BU) monotherapy. This study furthers the understanding of cell kinetics in CML. IFN therapy induces apoptosis and suppresses cell proliferation. The rate of programmed cell death prior to therapy and the extent of IFN‐triggered apoptosis exert a significant predictive impact on survival. In this study, ISEL‐positive (apoptotic) cells and bodies do not correspond to unscheduled cell repair as detected by PCNA immunoreactivity. © 1997 John Wiley & Sons, Ltd.

Gasless videoendoscopic implantation of aortobifemoral vascular prostheses in animal subjects is both practicable and reproducible using either a transperitoneal or extraperitoneal approach. The extraperitoneal approach has proven advantages over the transperitoneal approach. The in vitro fluid tightness achieved with endoscopically sutured aortic end-to-side anastomoses is comparable to conventionally sutured anastomoses.

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The gasless videoendoscopic implantation of GELSOFT aortobifemoral vascular prostheses times 6×6 mm in diameter using an extraperitoneal approach was tested in ten porcine experimental models at the Surgical Department of the University of Cologne, Germany. Gasless videoendoscopic surgery is performed with a laparolift-laparofan system. Aortobifemoral GELSOFT prostheses were successfully implanted in nine of ten animals, whereby one animal died during preparations for surgery of massive coronary infarctions. Average surgical durations using the extraperitoneal approach were 270 min. Dissection of the infrarenal aorta until occlusion took 45 min, average aortic occlusion 75 min, and iliacofemoral occlusion 45 min for the left side and 75 min for the right side. After successful videoendoscopic implantation of aortobifemoral GELSOFT prostheses all nine animals underwent laparotomy and resection of the aortobifemoral prosthetic segment. The quality of the endoscopically sutured aortic end-to-side anastomoses was examined in vitro under artificial circulation of glycerol/Ringer's lactate solution for evaluation of possible leakage and bursting pressures and then compared to conventionally sutured end-to-side anastomoses of 6-h-old porcine abdominal aorta and GELSOFT prostheses 6 mm in diameter. The maximum bursting pressure of all endoscopically sutured anastomoses was 480 mmHg mean pressure; the minimum was 140 mmHg mean pressure. The minimum leakage per minute was less than 10 ml/min for systolic pressure values between 120 and 350 mmHg. All endoscopically sutured aortic end-to-side anastomoses were comparable to conventionally sutured anastomoses concerning in vitro evaluation of bursting pressure and leakage per minute.