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Nuclear instruments & methods in physics research / A 1039, 167046 (2022). doi:10.1016/j.nima.2022.167046, Published by North-Holland Publ. Co., Amsterdam

In January 2018 the recent revision of the S2k guidelines on calculated parenteral initial treatment of bacterial diseases in adults-update 2018 (Editor: Paul Ehrlich Society for Chemotherapy, PEG) was realized. It is a helpful tool for the complex infectious disease setting in an intensive care unit. The present summary of the guidelines focuses on the topics of anti-infective agents, including new substances, pharmacokinetics and pharmacodynamics as well as on microbiology, resistance development and recommendations for calculated drug therapy in septic patients. As in past revisions the recent resistance situation and results of new clinical studies are considered and anti-infective agents are summarized in a table.

Sepsis-induced changes in pharmacokinetic parameters are a well-known problem in intensive care medicine. Dosing of antibiotics in this setting is therefore challenging. Alterations to the substance-specific kinetics of anti-infective substances have an effect on the distribution and excretion processes in the body. Increased clearance and an increased distribution volume (Vd) and particularly compromized organ function with reduced antibiotic elimination are often encountered in patients with sepsis. Renal replacement treatment, which is frequently used in intensive care medicine, represents a substantial intervention in this system. Current international guidelines recommend individualized dosing strategies and adaptation of doses according to measured serum levels and pharmacokinetic/pharmacodynamic (PK/PD) parameters as concepts to optimize anti-infective therapy in the critically ill. Likewise, the recommendation to adjust the administration form of beta-lactam antibiotics to prolonged or continuous infusion can be found increasingly more often in the literature. This article reviews the background of the individual dosing in intensive care patients and their applicability to the clinical routine.

The mortality of patients with sepsis and septic shock is still unacceptably high. An effective antibiotic treatment within 1 h of recognition of sepsis is an important target of sepsis treatment. Delays lead to an increase in mortality;therefore, structured treatment concepts form a rational foundation, taking relevant diagnostic and treatment steps into consideration. In addition to the assumed focus and individual risks of each patient, local resistance patterns and specific problem pathogens must be taken into account for selection of anti-infection treatment. Many pathophysiological alterations influence the pharmacokinetics of antibiotics during sepsis. The principle of standard dosing should be abandoned and replaced by an individual treatment approach with stronger weighting of the pharmacokinetics/pharmacodynamics (PK/PD) index of the substance groups. Although this is not yet the clinical standard, prolonged (or continuous) infusion of beta-lactam antibiotics and therapeutic drug monitoring (TDM) can help to achieve defined PK targets. Prolonged infusion is sufficient without TDM but for continuous infusion TDM is basically necessary. A further argument for individual PK/PD-oriented antibiotic approaches is the increasing number of infections due to multidrug resistant pathogens (MDR) in the intensive care unit. For effective treatment antibiotic stewardship teams (ABS team) are becoming more established. Interdisciplinary cooperation of the ABS team with infectiologists, microbiologists and clinical pharmacists leads not only to a rational administration of antibiotics but also has a positive influence on the outcome. The gold standards for pathogen detection are still culture-based detection and microbiological resistance testing for the various antibiotic groups. Despite the rapid investigation time, novel polymerase chain reaction (PCR)-based procedures for pathogen identification and resistance determination, are currently only an adjunct to routine sepsis diagnostics due to the limited number of studies, high costs and limited availability. In complicated septic courses with multiple anti-infective treatment or recurrent sepsis, PCR-based procedures can be used in addition to therapy monitoring and diagnostics. Novel antibiotics represent potent alternatives in the treatment of MDR infections. Due to the often defined spectrum of pathogens and the practically absent resistance, they are suitable for targeted treatment of severe MDR infections (therapy escalation).

Low Gain Avalanche Detector (LGAD) technology has been used to design and construct prototypes of time-zero detector for experiments utilizing proton and pion beams with High Acceptance Di-Electron Spectrometer (HADES) at GSI Darmstadt, Germany. LGAD properties have been studied with proton beams at the COoler SYnchrotron (COSY) facility in J\"ulich, Germany. We have demonstrated that systems based on a prototype LGAD operated at room temperature and equipped with leading-edge discriminators reach a time precision below 50 ps. The application in the HADES, experimental conditions, as well as the test results obtained with proton beams are presented., Comment: published in Eur. Phys. J. A 56, 183 (2020)

The purpose of this study was to identify thrombin-activatable fibrinolysis inhibitor (TAFI) in saliva and to investigate the correlation between TAFI levels in saliva and plasma. Subjects included were healthy adults without diseases or medication that could affect coagulation. Samples of stimulated saliva and blood samples were obtained from 33 subjects. Levels of TAFI in saliva and plasma were analysed. The association between levels of TAFI in saliva and plasma was calculated using linear regression. Low levels of TAFIa/TAFIai were found in most saliva samples but only one sample had levels that were above the lower limit of detection of the assay used. TAFI (proenzyme) was not found in saliva, so no correlations could be calculated. In this study there was no indication that there is TAFI present in secreted saliva. Either TAFIa/TAFIai in saliva were much lower than in plasma and under the detection limit of the assay used, or there was no TAFIa/TAFIai in the saliva tested.

Simulations of the electrophysiological behavior of the heart improve the comprehension of the mechanisms of the cardiovascular system. Furthermore, the mathematical modeling will support diagnosis and therapy of patients suffering from heart diseases. In this paper, the chain of modeling of the electrical function in the heart is described. The components are explained briefly, namely modeling of cardiac geometry, reconstructing the cardiac electrophysiology and excitation propagation. Additionally, the mathematical methods allowing to implement and solve these models are outlined. The three recently more investigated cases atrial fibrillation, ischemia and long-QT syndrome are described and show how cardiac modeling can support cardiologists in answering their open questions.

The 1st revision of the S2k guideline on the prevention and follow-up care of sepsis, provided by the German Sepsis Society in collaboration with 17 German medical scientific societies and one self-help group provides state-of-the-art information on the effective and appropriate medical care of critically ill patients with severe sepsis or septic shock. The guideline recommendations may not be applied under all circumstances. It rests with the clinician to decide whether a certain recommendation should be adopted or not, taking into consideration the unique set of clinical facts presented in connection with each individual patient as well as the available resources.

Five patients (four males and one female, aged 29 to 75 years) with subaortic stenoses were examined by transthoracal and transoesophageal echocardiography, including two-dimensional Doppler echocardiography. In only one patient could the obstruction of the left ventricular outflow tract be definitively evaluated by transthoracal echocardiography, whereas by transoesophageal echocardiography the presence of a subaortic membrane could be demonstrated in all patients. The latter method thus proved to be reliable in evaluating the extent of obstruction.

Die Verknupfung der Vorschriften der Trinkwasserverordnung (TrinkwV) 2001 mit dem umfangreichen Technischen Regelwerk sowie mit den Empfehlungen des Umweltbundesamtes (UBA) und der Trinkwasserkommission hat sich prinzipiell bewahrt. Die TrinkwV 2001 beinhaltet aber auch einige Passagen, die eine Novellierung angezeigt erscheinen lassen. Vom Verordnungsgeber wird an ihre Uberarbeitung gedacht. Dabei werden eigene Erkenntnisse sowie Erfahrungen der Lander und der Wasserversorger beim Vollzug der Verordnung zu berucksichtigen sein. Es ist vorgesehen, Verbesserungen sowie weitere Vereinfachungen zum Burokratieabbau vorzunehmen, ohne dabei vom existierenden hohen Schutzniveau fur das Trinkwasser in Deutschland abzuweichen.

We report the results of 84 patients with ALL after related (n = 46) or unrelated (n = 38) allogeneic SCT. Mean recipient age was 23 years (range: 1-60) and median follow-up was 18 months (range: 1-133). Forty-three patients were transplanted in CR1; 25 in CR2 or CR3; four were primary refractory; four in PR; eight in relapse. The conditioning regimen consisted of TBI/VP16/CY (n = 76), TBI/VP16 (n = 2), TBI/CY (n = 2), Bu/VP16/CY (n = 4). The OS at 3 years was 45% (44% unrelated, 46% related). Univariate analysis showed a significantly better OS for patients

Combined laser and synchrotron radiation (SR) photoelectron spectroscopy with a temporal resolution of 60 ps has been applied to study the relaxation of the surface photovoltage (SPV) after laser pulse excitation for a SiO 2 /Si(1 0 0) interface. For zero time delay between laser and SR pulses the Si 2p core level exhibits an initial spectral shift of 300 meV which relaxes non-exponentially to about 100 meV after 800 ns. The relaxation of the SPV can be modeled using a self-decelerating carrier relaxation based on a thermionic emission model.

We investigated toxicity and efficacy of in vivo T-cell depletion with anti-thymocyte globulin (ATG) as part of an intensified myeloablative conditioning regimen followed by allogeneic stem cell transplantation in patients with advanced multiple myeloma. The conditioning regimen consisted of modified total body irradiation, busulfan and cyclophosphamide (n=15) or in the case of prior dose-limiting radiotherapy of busulfan and cyclophosphamide (n=3). The median age was 44 years (range, 29-53) and the median time from diagnosis to transplant was 12 months (range, 6-144). Grade II-IV acute graft-versus-host disease (GvHD) occurred in six patients (35%). Severe grade III/IV GvHD developed in one patient (6%). Three patients died of therapy-related causes (17%). A complete remission (CR) with negative immunofixation after allogeneic transplantation was seen in eight of the evaluable patients (53%). After a median follow-up of 41 months (range, 8-84), the estimated overall survival at 6 years for all patients is 77% (CI 95%: 58-96%). The estimated progression-free survival at 6 years for all patients is 31% (CI 95%: 2-59%) and 46% (CI 95%: 9-83%) for patients with CR. In vivo T-cell depletion with ATG resulted in a low rate of severe GvHD with low treatment-related mortality, and a substantial number of long-term survivors.

Der bisher in der Bemessungspraxis wegen seiner Einfachheit bevorzugte Nachweis der Biegeschlankheit als Ersatz einer komplizierten Durchbiegungsberechnung kann nach den vorliegenden Untersuchungsergebnissen mit einigen Modifikationen weiterhin als ausreichend und ingenieurmasig angesehen werden. Zur Erfassung des Einflusses der Bauteilgeometrie auf die Bauteilverformungen wird fur die Anwendung in der Praxis ein Naherungsansatz vorgeschlagen, bei dem zwischen Balken- und Plattentragwerken unterschieden wird.

Die nachfolgenden Darlegungen zeigen die Ergebnisse der baustofflichen und statisch-konstruktiven Untersuchungen zur Entwicklung von neuartigen Plattenelementen aus Polymerbeton. Hierbei handelt es sich um eine Gemeinschaftsarbeit des Fachbereiches Bauingenieurwesen der Universitat Rostock und dem Mineralguszentrum Neubukow. Das Ziel der baustofflichen Betrachtungen bestand in der kennwertorientierten Analyse des Werkstoffes Polymerbeton. Dazu gehorte die Optimierung der Mischung aus einem polymeren Bindemittel mit einem mineralischen Zuschlag sowie die Ermittlung mechanischer temperaturabhangiger Festbetonkenngrosen. Im Rahmen der statisch-konstruktiven Untersuchungen wurde das Tragverhalten von Plattenstreifen unter Kurzzeit- und Dauerlast untersucht. Die so gewonnenen Ergebnisse bildeten die Grundlage zur Erarbeitung eines Nachweiskonzeptes. Das Ergebnis ist eine 3,5 cm dicke dauerbestandige, oberflachenfertige sowie vermarktungsfahige Balkonbodenplatte.

One-hundred and two patients with good risk myeloid leukemia (CML first chronic phase or AML first CR) were transplanted from HLA-related donors after conditioning with (n = 45) or without anti-thymocyte globulin (ATG) (n = 57). One graft failure was observed in the non-ATG and none in the ATG group. The median time to leukocyte engraftment (> 1 x 10(9)/l) was 16 (range 12-33) in the ATG group and 17 days (range 11-29) in the non-ATG group (NS) and for platelet engraftment (> 20 x 10(9)/l) 24 and 19 days (P = 0.002), respectively. Acute GVHD grade II-IV was observed in 47% of the non-ATG and in 20% of the ATG group (P = 0.004). Grade III/IV GVHD occurred in 7% of the ATG and in 32% of the non-ATG group (P = 0.002). Chronic GVHD was seen in 36% and 67% (P = 0.005), respectively. After a median follow-up of 48 months (range 2-128), the 5-year estimated OS is 66% (95% KI: 51-81%) for the ATG group and 59% (95% KI: 46-72%) for the non-ATG group (NS). The 5-year estimated DFS is 64% (95% KI: 50-78%) for ATG and 55% (95% KI: 43-67%) for the non-ATG regimen (NS). The 5-year probability of relapse was 5% in the ATG and 15% in the non-ATG group (NS). ATG as part of the conditioning regimen leads to a significant reduction in GVHD without increase of relapse in patients with myeloid leukemia after stem cell transplantation from HLA-related donors.

A major problem after autologous or allogeneic stem cell transplantation is prolonged thrombocytopenia. There are several studies published about correlations of the composition of the graft and time to platelet engraftment for autologous transplantation but only a few studies for allogeneic transplantation. In our study, we wanted to find out whether the correlation between the time to platelet engraftment and amount of transplanted CD34(+)CD41(+) cells described previously after autologous peripheral blood stem cell transplantation could be reproduced in the allogeneic bone marrow transplantation setting. We found correlations not only for the number of transplanted CD34(+) cells with the time to leukocyte engraftment (r = -0.32, p = 0.045) but also for the number of transplanted CD34(+)CD41(+) cells and time to platelet engraftment (r = -0.34, p = 0.038), which were both statistically significant. A significant correlation between transplanted CD34(+) cells versus platelet engraftment and transplanted CD34(+)CD41(+) cells versus leukocyte engraftment was not found. The finding that the amount of committed megakaryocyte progenitor cells in the graft is an important predictive factor for platelet engraftment after allogeneic bone marrow transplantation might be the base for future studies of ex vivo expansion of clonable megakaryocyte precursors.

A device for quantitative morphometric analysis of tissue alterations caused by laser or electrosurgery was developed. The unit was designed for interactive evaluation of histological specimens obtained by experimental surgery. It consists of a microscope, a stepper-motor driven scanning stage, a digitizing tablet and a PC with software developed for this purpose. The depth of the incision, the width and the area of the coagulation zone can be measured independent of the microscopic magnification. Graphs and data can be exported in standard data-formats (ASCII, HPGL, DXF) for further processing. The application is demonstrated on experimental incisions with a CO2-laser in the oral mucosa of pigs; quantitative data are presented.

We investigated the feasibility and efficacy of a fludarabine-based dose-reduced conditioning regimen followed by stem cell transplantation from related (n = 5) or unrelated HLA-matched donors (n = 7) in 12 patients with high risk MDS, who were not eligible for a standard myeloablative conditioning regimen. The conditioning regimen consisted of fludarabine 30 mg/m(2) daily for 6 days, busulfan 4 mg/kg daily for 2 days and anti-thymocyte globulin (ATG, rabbit) 10 mg/kg daily for 4 days in 11 patients, while one patient received fludarabine, ATG, cyclophosphamide and thiotepa. Graft-versus-host disease prophylaxis consisted of cyclosporine and a short course of methotrexate. The median age of the patients was 53 years (range 37-59). The median percentage of blasts in bone marrow aspirate at transplantation was 15% (range

Langjahrig unterlassene Kalkung fuhrte bei mineralisch (NPK) und mit Stallmist gedungtem Boden in den letzten 20 Versuchsjahren zu pH‐Werten von 4,2 bis 3,6. Die Relativertrage nahmen im Vergleich zur mineralischen Volldungung (NPKCa) ab. Diese Abnahme war von der Pflanzenart abhangig und bei Sommergerste deutlicher als bei Kartoffeln und Mais. Kalkung des mit NPK gedungten Bodens erhohte die Ertrage bei Mais bereits im ersten Jahr, bei Gerste und Kartoffeln erst nach mehreren Jahren auf das Niveau der Vergleichsvariante. Bei einer Umstellung von Stallmist auf Stallmist + NPKCa glichen sich die Ertrage von Mais und Sommergerste im gleichen Jahr, bei Kartoffeln erst nach mehreren Jahren dem langjahrig mit Stallmist + NPKCa gedungten Prufglied an. Im langjahrig ungedungten Boden fielen pH‐Wert, KDL und PDL in den letzten 20 Versuchsjahren ebenfalls deutlich ab (pH 3,6 bis 4,1; KDL 2,2 bis 5,0 und PDL 3,5 bis 5,5 mg/100 g Boden). In deren Folge und durch N‐Mangel kam es bei Gerste zum Totalausfall und bei Ma...

To evaluate the efficacy and toxicity of two different etoposide (VP-16) dosages (30 or 45 mg/kg) in combination with busulfan/cyclophosphamide as conditioning therapy followed by stem cell transplantation in acute myeloid leukemia (AML), 90 patients with AML received either 30 mg/kg (n = 60) or 45 mg/kg (n = 30) etoposide in combination with busulfan (16 mg/kg) and cyclophosphamide (120 mg/kg). The stem cell source was allogeneic related bone marrow (BM) (n = 53), allogeneic unrelated BM (n = 5), allogeneic unrelated peripheral blood (PBSC) (n = 2), syngeneic BM (n = 2), autologous BM purged (n = 9) or unpurged (n = 9), autologous PBSC (n = 10). Fifty-six patients (62%) were in first CR, 26 (29%) were > first CR, and eight (9%) were transplanted in relapse. Principal toxicities in both groups were mucositis and hepatotoxicity. Forty-five mg/kg etoposide resulted in greater hepatic toxicity (P = 0.03), and a higher incidence of VOD (23 vs 12%, P = 0.04) and acute GVHD grade III/IV (13 vs 5%, NS). The treatment-related mortality was 17% in the 30 mg/kg group and 33% in the 45 mg/kg group, mainly due to infections, intestinal pneumonia and GVHD. Hematological recovery of leukocytes 1/nl was comparable in both groups (17 vs 16 days). After a median follow-up of 16 months 19% in the 30 mg/kg group and 23% in the 45 mg/kg group relapsed. In patients who had undergone allogeneic related bone marrow transplantation in first CR no relapses occurred after a median follow-up of 3 years. For all patients the 3-year estimated disease-free survival was 62% in the 30 mg/kg group and 40% in the 45 mg/kg group (P = 0.03). For patients in first CR who underwent allogeneic related stem cell transplantation the 3 year disease-free survivals were 80% and 66%, respectively (P = 0.4). We conclude that etoposide 30 mg/kg or 45 mg/kg in combination with busulfan/cyclophosphamide is a highly active regimen for bone marrow transplantation of patients with AML with a low relapse rate. However, conditioning with 30 mg/kg rather than 45 mg/kg etoposide resulted in less toxicity and a better overall survival due to a lower transplant-related mortality. Bone Marrow Transplantation (2000) 26, 711-716.

At the time of primary therapy (surgery, systemic chemotherapy and/or radiation), disseminated tumor cells in the bone marrow can be found in almost one-third of patients with cancer of the breast, ovary, esophagus, stomach, colon, and other solid tumors. Whereas the prognostic impact of the mere presence of these cells is still a matter of debate, it has been shown that expression of tumor-associated antigens in disseminated tumor cells is linked to more aggressive disease. Therefore, further characterization of disseminated tumor cells at the protein and gene level has become increasingly important. To date, the most common detection method for disseminated tumor cells in the bone marrow is an immunocytochemical approach using cytokeratin-directed antibodies for detection of epithelial cells and the APAAP system for their visualization. We have established a new double immunofluorescence technique enabling simultaneous detection, phenotyping, and antigen quantification of disseminated tumor cells. Mononuclear cells from bone marrow are enriched by Ficoll gradient centrifugation and cytospins are prepared. Double immunofluorescence is performed using antibodies against cytokeratins 8/18/19 (mAb A45B/B3) and the uPA receptor CD87 (pAb HU277). CD87 expression is recorded by confocal laser scanning microscopy (CLSM) using fluorescence labeled latex beads as the reference; staining intensities of all the scans are then summed and quantified (extended focus). This protocol, originally designed for disseminated tumor cells in bone marrow, can also be applied to disseminated tumor cells in blood, to leukapheresis cells or to cells present in malignant ascites or other malignant effusions. The tumor cells detected may be used for gene and mRNA analyses. Furthermore, disseminated tumor cells also represent interesting targets for clinical studies on patient prognosis or prediction of therapy response as well as for specific tumor-biological therapies.

The reverse transcription polymerase chain reaction (RT-PCR) amplification of cytokeratin 20 (CK20) mRNA is considered a promising candidate method for the detection of circulating tumour cells in bone marrow and peripheral blood of cancer patients. In this study we have investigated the diagnostic specificity of the CK20 mRNA detection in samples from healthy donors (HD; n = 33), intensive care units patients (ICU; n = 20) and bone marrow obtained from patients suffering from chronic inflammatory diseases (CID; n = 14). RNAs purified from stabilized lysates showed positive results in 24% of the HD group (8/33), 35% of the ICU group (8/20) and in 40% of the CID group (5/14). The use of Ficoll gradients to separate nucleated cells completely restored the specificity of this CK20 RT-PCR assay. The CK20-expressing cells are positively identified to belong to the granulocyte fraction of leucocytes, which appear to express the gene on a background level. Our results demonstrate for the first time that CK20 mRNA expression is not limited to epithelium. Its occurrence in normal granulocytes has to be considered in tests designed to detect circulating cancer cells or micrometastases. © 1999 Cancer Research Campaign

Infections are a major cause of morbidity and mortality in patients undergoing high-dose therapy and subsequent autologous or allogeneic haemopoietic stem cell transplantation, despite the change from topical to systemic anti-infection prophylaxis and the introduction of growth factors and new antimicrobial drugs. We report our single centre experience with data from 409 patients treated at our unit from its opening in 1990 until May 1997. Three hundred and seventy-eight patients were transplanted for the first time, 12 patients were retransplanted or boosted and 19 patients were readmitted for miscellaneous reasons. 245 patients were allografted and 157 autografted. Antimicrobial prophylaxis was mainly quinolones, fluconazole plus amphotericin-B orally, aciclovir, and TMP/SMX or pentamidine. Three hundred and nineteen (78%) developed fever of significantly longer duration in the allogeneic setting with anti-CMV seropositivity. The most frequent infection was fever of unknown origin (50.6%), followed by septicaemia (12.5%) and pneumonia (11.0%). Pathogens isolated in 24.6% of the infections were mostly gram-positive bacteria (57.9%), followed by non-fermenting rods (11.2%), Aspergillus spp. and Candida spp. (10.3%, each). Cumulative response rate to antimicrobial therapy was 66.9%. Infections were responsible for 62.5% (25/40) of deaths after transplantation. Predominant pathogens were Aspergillus spp. (11), Candida spp. (four), and Pseudomonas spp. (three). None of the patients died from gram-positive bacterial infection. The risk of dying from infection was 11.2% after allografting and 0.8% after autotransplantation. Infections remain a major risk for early death after allogeneic transplantation of haemopoietic stem cells. Infection with gram-negative bacteria can be prevented by quinolone prophylaxis. Predominant pathogens are Aspergillus spp. Candida spp. and nonfermenting rods. Systemic infection with these pathogens is associated with a poor prognosis. Antimycotic prophylaxis and the therapy must be improved.

Cytokines are involved in hematopoiesis and cell growth regulation. Many cytokines have the property of increasing the number of hematopoietic progenitor cells circulating in peripheral blood. Since it has been shown that peripheral blood progenitor support after myeloablative therapy resulted in a faster engraftment than after autologous bone marrow support, recent effort is focusing on optimizing stem cell harvest by different cytokines, cytokine combination, or evaluation of dose and schedule dependencies. One of the first cytokines shown to mobilize stem cells within peripheral blood was the granulocyte-stimulating factor (G-CSF). This review is focusing on the current state of cytokine-induced mobilization of stem cells in patients with hematological malignancies or solid tumors.

High-dose therapy with subsequent allogeneic or autologous transplantation of haemopoietic progenitor cells has proved to be an important therapy for patients with haematological malignancies, certain non-haematological cancers and several nonmalignant disorders such as severe aplastic anaemia. Allogeneic and autologous transplantation are associated with specific complications. One major problem after allotransplantation is the graft versus host disease with jaundice, diarrhoea and erythema. The major problem of autologous stem cell transplantation is a potential reinfusion of contaminating tumor cells. Allogeneic transplantation has been refined, and an approach to modulate graft versus host disease is a partial or complete lymphocyte depletion. Especially in the allogeneic setting, the field of cell therapy for modulation of graft versus host disease and treatment of transplant-related complications is expanding. For autologous transplantation different techniques to purge autografts from malignant contaminants have been developed. These cell selection techniques consist of immunological, biological, cultural, chemotherapeutical and physical techniques. This article reviews techniques currently used for graft engineering in bone marrow and blood stem cell transplantation.

We investigated an intensified conditioning regimen including fractionated total body irradiation (12 Gy), etoposide (30–45 mg/kg) and cyclophosphamide (120 mg/kg), followed by autologous (n = 5), allo-related (n = 13) or allo-unrelated (n = 6) bone marrow (n = 22) or peripheral stem cell (n = 2) transplantation in patients with Philadelphia chromosome-positive acute lymphoblastic leukemia. One patient received busulfan (16 mg/kg) instead of TBI. Nineteen patients were transplanted in 1CR, two in 2CR, one in 1PR and two in relapse. Major toxicity was mucositis grade II according to the Bearman scale in all patients. The treatment-related mortality was 25%, mainly due to infection or GVHD after allogeneic transplantation. After a median follow-up of 45 months (range 2–93), nine patients (37.5%) remain alive in CR. Nine patients (37.5%) relapsed and eight (33.3%) of these subsequently died. After autologous transplantation, four of five patients (80%) relapsed and died. Late relapse was seen after allogeneic, as well as autologous transplantation, at 33 and 59 months, respectively. The Kaplan–Meier estimate of leukemia-free survival for all patients is 38% at 3 years (95% CI: 18–58%) and 35% at 5 years (95% CI: 15–55%). For allogeneic transplants in first CR (n = 15) the estimate of disease-free survival was 46% at 3 years (95% CI: 19–73%) and 34% at 5 years (95% CI: 17–51%). Patients aged below 30 years had a better estimated overall survival at 3 years (61% vs 11%, P

Several reverse transcriptase polymerase chain reaction (RT-PCR) assays have been described for the detection of circulating tumour cells in blood and bone marrow. Target mRNA sequences for this purpose are the cytokeratins (CK) 19 and 20, the carcinoembryonic antigen (CEA), and the prostate-specific antigen messages. In this study, we investigated biological factors influencing the specificity of the CK19 and CEA RT-PCR assays. Bone marrow, granulocyte colony-stimulating factor (G-CSF)-mobilized blood stem cells and peripheral blood samples obtained from healthy volunteers (n = 15; CEA n = 7), from patients with epithelial (n = 29) and haematological (n = 23) cancer and from patients with chronic inflammatory diseases (n = 16) were examined. Neither CEA nor cytokeratin 19 messages could be amplified from bone marrow samples from healthy subjects and from patients with haematological malignancies. In contrast, specimens from patients with inflammatory diseases scored positive up to 60%. To investigate the influence of inflammation on target mRNA expression, haemopoietic cells were cultured with and without cytokine stimulation in vitro. CK19 messages could be easily detected in cultured marrow cells without further stimulation, CEA messages only after gamma-interferon (gamma-INF) stimulation. In contrast, G-CSF-mobilized peripheral blood stem cells were positive for CK19 messages only after stem cell factor (SCF) or interleukin stimulation. We conclude that transcription of so-called tissue-specific genes is inductible in haemopoietic tissues under certain conditions. These factors have to be considered in future applications of RT-PCR for the detection of minimal residual disease.