Persistence of pre-leukemic clones during first remission and risk of relapse in acute myeloid leukemia.

Some patients with acute myeloid leukemia (AML) who are in complete remission after induction chemotherapy harbor persisting pre-leukemic clones, carrying a subset of leukemia-associated somatic mutations. There is conflicting evidence on the prognostic relevance of these clones for AML relapse. Here, we characterized paired pre-treatment and remission samples from 126 AML patients for mutations in 68 leukemia-associated genes. Fifty patients (40%) retained ⩾1 mutation during remission at a variant allele frequency of ⩾2%. Mutation persistence was most frequent in DNMT3A (65% of patients with mutations at diagnosis), SRSF2 (64%), TET2 (55%), and ASXL1 (46%), and significantly associated with older age (P<0.0001) and, in multivariate analyses adjusting for age, genetic risk, and allogeneic transplantation, with inferior relapse-free survival (hazard ratio, 2.34; P=0039) and overall survival (hazard ratio, 2.14; P=036). Patients with persisting mutations had a higher cumulative incidence of relapse before, but not after allogeneic stem cell transplantation. Our work underlines the relevance of mutation persistence during first remission as a novel risk factor in AML. Persistence of pre-leukemic clones may contribute to the inferior outcome of elderly AML patients. Allogeneic transplantation abrogated the increased relapse risk associated with persisting pre-leukemic clones, suggesting that mutation persistence may guide postremission treatment.Leukemia accepted article preview online, 18 December 2017. doi:10.1038/leu.2017.350.

Early assessment of minimal residual disease in AML by flow cytometry during aplasia identifies patients at increased risk of relapse.

In acute myeloid leukemia (AML), assessment of minimal residual disease (MRD) by flow cytometry (flow MRD) after induction and consolidation therapy has been shown to provide independent prognostic information. However, data on the value of earlier flow MRD assessment are lacking. Therefore, the value of flow MRD detection was determined during aplasia in 178 patients achieving complete remission after treatment according to AMLCG (AML Cooperative Group) induction protocols. Flow MRD positivity during aplasia predicted poor outcome (5-year relapse-free survival (RFS) 16% vs 43%, P<0.001) independently from age and cytogenetic risk group (hazard ratio for MRD positivity 1.71; P=0.009). Importantly, the prognosis of patients without detectable MRD was neither impacted by morphological blast count during aplasia nor by MRD status postinduction. Early flow MRD was also evaluated in the context of existing risk factors. Flow MRD was prognostic within the intermediate cytogenetic risk group (5-year RFS 15% vs 37%, P=0.016) as well as for patients with normal karyotype and NPM1 mutations (5-year RFS 13% vs 49%, P=0.02) or FLT3-ITD (3-year RFS rates 9% vs 44%, P=0.016). Early flow MRD assessment can improve current risk stratification approaches by prediction of RFS in AML and might facilitate adaptation of postremission therapy for patients at high risk of relapse.

Sequential therapy combining clofarabine and T-cell-replete HLA-haploidentical haematopoietic SCT is feasible and shows efficacy in the treatment of refractory or relapsed aggressive lymphoma.

Prognosis is poor for patients with biologically aggressive Non-Hodgkin lymphoma (NHL), refractory to chemotherapy or relapsed after autologous transplantation, especially when no disease control before allogeneic transplantation is achieved. In 16 patients (median age 53, median prior regimes 5) with relapsed or refractory non-remission NHL, we analysed retrospectively the efficacy of a sequential therapy comprising clofarabine re-induction followed by a reduced-intensity conditioning with fludarabine, CY and melphalan, and T-cell-replete HLA-haploidentical transplantation. High-dose CY was utilized post-transplantation. All patients engrafted. Early response (day +30) was achieved in 94%. Treatment-related grade III-IV toxicity occurred in 56%, most commonly transient elevation of transaminases (36%), while there was a low incidence of infections (19% CMV reactivation, 19% invasive fungal infection) and GVHD (GVHD: acute III-IV: 6%; mild chronic: 25%). One-year non-relapse mortality was 19%. After a median follow-up of 21 months, estimated 1- and 2-year PFS was 56 and 50%, respectively, with 11 patients (69%) still alive after 2 years. In summary, sequential therapy is feasible and effective and provides an acceptable toxicity profile in high-risk non-remission NHL. Presumably, cytotoxic reinduction with clofarabine provides enough remission time for the graft-versus lymphoma effect of HLA-haploidentical transplantation to kick in, even in lymphomas that are otherwise chemo-refractory.

Lack of association between childhood stroke after varicella and human leukocyte antigen (HLA)-B51.

Human leukocyte antigen (HLA)-B51 has been suggested as an immunogenetic marker for a genetic predisposition to vascular occlusion in response to an immunological stimulus. Varicella has been reported to be a possible risk factor for stroke. We performed DNA-based HLA typing in 11 young patients (mean age: 5.2 years) with unexplained ischaemic stroke. In eight of them varicella had occurred before their stroke. HLA-B51 was negative in all 11 patients and we did not find any significant accumulation of other HLA-subgroups. Our study does not support an association between susceptibility to stroke after varicella and HLA-B51.

Simultaneous quantification of citalopram, clozapine, fluoxetine, norfluoxetine, maprotiline, desmethylmaprotiline and trazodone in human serum by HPLC analysis.

We describe an analytical procedure for the simultaneous quantification of citalopram (seropram), clozapine (leponex), fluoxetine (fluctine), norfluoxetine, maprotiline (ludiomil), desmethylmaprotiline and trazodone (trittico) in human serum within a period of 11.5 minutes using reversed phase HPLC. After 2 liquid/liquid extractions in the sample preparation phase, the drugs and metabolites were separated on a C18 column using a mobile phase consisting of acetonitrile/buffer (30/70, v:v) at 70 degrees C, a flow rate of 1.5 m/min and haloperidol as internal standard. Absorption and native fluorescence signals of the eluted compounds were detected simultaneously at 260 nm and 227/300 nm (excitation/emission), respectively. The calibration ranges for citalopram, clozapine, fluoxetine, norfluoxetine, maprotiline, and desmethylmaprotiline ranged from 50-400 microg/l and for trazodone from 50-3,200 microg/l. The CVs varied between 0.6% and 5.5% (within-run) and between 3.2% and 7.1% (between-run). Recoveries were > 90% for all pharmaceuticals. We noticed no interferences from several commonly used drugs.

[Practical usage of a quality management system according to ISO 9001 : 2000 focusing on the double contrast (barium) enema]. / Praktische Anwendung eines Qualitätsmanagementsystems nach ISO 9001:2000 am Beispiel von Qualitätsverbesserungsmassnahmen beim Kolon-Kontrastmitteleinlauf, Germany.

PURPOSE: We describe the implementation of quality improvement measures in a quality management system. METHODS: With questionnaires for radiologists and patients, we investigated the relations between patient preparation and diagnostic quality serving the main purpose, to improve diagnostic quality with the help of more detailed patient information (e.g., changing preparation sheets and handling out "peri-med" information sheets to the patients). Furthermore, a comparative data ascertainment at other institutes was integrated. RESULTS: For the group of outpatients, increasing process quality (patient information) and outcome (diagnostic quality) could be achieved. Taking aspects of quality costs into consideration, a decrease in costs due to failures was achieved. CONCLUSION: More detailed patient information has positive effects on diagnostic quality of the double contrast (barium) enema.

[Titanium nitride cardiac pacemaker electrodes]. / Titannitrid-Herzschrittmacher-Elektroden.

The sensing and pacing performance of pacemaker electrodes is characterized by the electrochemical properties of the electrodes/tissue layer; the usually smooth metallic electrode surface results in a high pass filter characteristic. Consequently, the detected intracardiac signals, which control the implantable systems, are not optimally matched to the spectral contents of the depolarisation signal. To avoid interference caused by noise (EMI, muscle potentials, etc.) a shift of the frequency of the band pass towards the lower frequency spectrum is required. As previously reported, the electrochemical properties of sintered and surface-treated electrodes prove the predicted improvement of sensing performance if titanium-nitride coated electrodes are used. Our results demonstrate their superiority above all the other electrodes presently known. The advantages can be referred to the micro-crystalline surface structure achieved by sputter-deposited electrode coatings and the kinetics of the ionic exchange. Furthermore, the acute thresholds achieved with the TiN-systems were significantly better than those of the smooth metallic surface. These results were also confirmed for chronic implants and are attributable to the known biocompatibility of titanium and its alloys.

[Motion energy as a control variable for adapting stimulation frequency]. / Bewegungsenergie als Steuergrösse für die Anpassung der Stimulationsfrequenz.

An activity sensing rate-responsive pacing system is presented which adaptively controls heart rate to adjust cardiac output in response to increased metabolic demand, and more optimally restore homeostasis of the intact cardiovascular system. The current use of ventricular demand and DDD universal pacing systems, although rate and multi-parameter and multi-function programmable, are fixed at these programmed settings. These devices are adequate for patients at rest or during moderate exertion, but are suboptimal for physically active patients whose physiology requires increased oxygen supply to meet an increased cardiac demand. In the past, these patients may have experienced fatigue or dyspnea out of proportion to their cardiovascular disease. The Ergos rate-adaptive single- and dual-chamber pacing system is a second generation pulse generator which is rate responsive to a patient's increased physiologic demand by sensing a motion signal which reflects increased work load and the need for a compensating increase in heart rate. Ergos offers increased assistance to patients with sinus bradycardia who may require the rate-responsiveness with the additional advantage of AV synchrony. Clinical results show the effectiveness of the presented sensor control by motion energy for rate adaptive pacing therapy.

[Analysis of myocardial evoked potentials in relation to ergometric load and heart rate for use in frequency-adaptive cardiac pacemakers]. / Analyse evozierter Myokardpotentiale in Abhängigkeit von ergometrischer Belastung und Schlagfrequenz für die Anwendung in frequenzadaptiven Herzschrittmachern.

The aim of this study was to evaluate the ventricular evoked response (VER) measured with unipolar fractally coated pacing leads for use with rate-responsive pacemakers. To this end, the morphology of the VER, its variation with pacing rate and various levels of physical loading, and the long-term stability of the signal were studied using the telemetric features of implantable pacemakers. Fractally coated electrodes were used in order to minimize the stimulation artefact, thus enabling a reliable unipolar measurement of the VER. The VER shows uniform basic morphology, and remains virtually unchanged after 3 months; moreover, frequency and load-dependent changes of VER morphology were identical for all patients. A special programming device has been developed to evaluate a rate adaptive algorithm for optimizing the pacing rate to the current loading situation, and was successfully tested in two patients. The algorithm was shown to be capable of calculating a heart rate adequate for haemodynamic demand.

Second haematopoietic SCT using HLA-haploidentical donors in patients with relapse of acute leukaemia after a first allogeneic transplantation.

Haploidentical haematopoietic SCT (HSCT) using T-cell-replete grafts and post-transplant high-dose CY has found increasing acceptance. Our purpose was to evaluate the feasibility and outcome of this strategy as second HSCT incorporating donor change for acute leukaemia relapse after a first allogeneic transplantation. The courses of 20 consecutive adults (median age 37 years, 12 male) with AML (n=14), ALL (n=5) and acute bi-phenotypic leukaemia (n=1) were analysed retrospectively. Conditioning consisted of fludarabine, CY and either melphalan or TBI or tresosulfan+/-etoposide. Engraftment was achieved in 17 (85%), and a second remission was induced in 15 patients (75%) on day +30. The rate of grade II-IV acute GvHD was 35%, while chronic GvHD occurred in five patients. Most commonly observed grade III-IV toxicities were mucositis (30%), hyperbilirubinemia (20%), elevation of transaminases (20%) and creatinine (20%), while invasive fungal infection affected 30%. One-year non-relapse mortality (NRM) was 36%. At a median follow-up of 17 months, estimated 1-year OS was 45%, and 1-year relapse-free survival was 33%. This strategy was feasible and allowed for successful engraftment with a moderate rate of toxicity. Early outcome and NRM are at least comparable with results after a second HSCT from HLA-matched donors without donor change at HSCT2.

Comparative molecular analysis of therapy-related and de novo acute promyelocytic leukemia.

Therapy-related acute promyelocytic leukemia (t-APL) has been reported as a late complication of exposure to radiotherapy and/or chemotherapeutic agents targeting DNA topoisomerase II. We have analyzed in t-APL novel gene mutations recently associated with myeloid disorders. Unlike previous reports in acute myeloid leukemia (AML), our results showed neither IDHs nor TET2 mutations in t-APL. However we found an R882H mutation in the DNMT3A gene in a patient with t-APL suggesting a possible role of this alteration in the pathogenesis of t-APL.

Monitoring of WT1 expression in PB and CD34(+) donor chimerism of BM predicts early relapse in AML and MDS patients after hematopoietic cell transplantation with reduced-intensity conditioning.

Relapse of malignant disease remains the major complication in patients with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS) after hematopoietic cell transplantation (HCT) with reduced-intensity conditioning (RIC). In this study, we investigated the predictive value of disease-specific markers (DSMs), donor chimerism (DC) analysis of unsorted (UDC) or CD34(+) sorted cells and Wilms' tumor gene 1 (WT1) expression. Eighty-eight patients with AML or MDS were monitored after allogenic HCT following 2 Gy total-body irradiation with (n=84) or without (n=4) fludarabine 3 × 30 mg/m(2), followed by cyclosporin A and mycophenolate mofetil. DSMs were determined by fluorescence in situ hybridization (FISH) and WT1 expression by real-time polymerase chain reaction. Chimerism analysis was performed on unsorted or CD34(+) sorted cells, by FISH or short tandem repeat polymerase chain reaction. Twenty-one (24%) patients relapsed within 4 months after HCT. UDC, CD34(+) DC and WT1 expression were each significant predictors of relapse with sensitivities ranging from 53 to 79% and specificities of 82-91%. Relapse within 28 days was excluded almost entirely on the basis of WT1 expression combined with CD34(+) DC kinetics. Monitoring of WT1 expression and CD34(+) DC predict relapse of AML and MDS after RIC-HCT.

A survey of contrast media used in coronary angiography.

In recent years, various contrast media have been developed for use in coronary angiography. These contrast media may be divided into ionic contrast media of high osmolality, those of low osmolality, and nonionic contrast materials. We conducted our own clinical studies with 40 patients. In random succession a standard contrast medium (ionic, of high osmolality) and a new-generation contrast medium (either nonionic or ionic with low osmolality) were injected into the right and left coronary arteries. After each injection we measured the systolic and diastolic blood pressure using a liquid-filled coronary catheter. In addition, the change in the length of the cardiac cycle was registered in terms of the R-R interval (in ms) and at the same time, leads I, II, and III of the ECG were recorded. We studied the influence of the various contrast media on the activity of ATPase in in vitro experiments, using Lasser and Lang's [30]. When ionic contrast media of low osmolality and nonionic contrast media were utilized the heart rate showed no change. Disturbances of rhythm such as ventricular tachycardia or fibrillation were not observed. All of the contrast media used produced the same ECG changes. These changes can be ascribed to the inhibition of ATPase. The arterial blood pressure was lowered significantly only by ionic contrast media of high osmolality only.

[Systolic periods in dependence of age]. / Die systolischen Zeitintervalle in Abhängigkeit vom Lebensalter.

In 278 healthy probands in the age of 23 to 80 years were measured the systolic periods and examined in dependence of age. There are found the following results: 1. from the youngest to the oldest group there is a small, statistically not significant increase of the middle systolic period. 2. the presphygmic phase (PEP) and the frequence-corrected interval (PEPI) show a small increase of the middle values in the first four age steps. Only after the age of 60 years is a clear increase of PEP and PEPI. 3. the expulsion time (LVET) and the expulsion index (LVETI) show in an increasing tendency of the mean values no significant difference of the mean value in the age groups. 4. the quotient LVET/PEP follows the course of PEP and PEPI. There is supposed, that the clear statistically significant increase of the mean value of the presphygmic phase and the presphygmic index after the age of 60 years cannot not be reduced to a "physiologic age insufficiency of the heart", but its cause is founded in asymptomatic courses of the coronary heart disease or other defined heart diseases.

[The resonance frequency of the radial artery in healthy untrained and endurance-trained older men]. / Resonanzfrequenz der Arteria radialis bei gesunden untrainierten und ausdauertrainierten älteren Männern.

There are examined 2 groups of two at a time men over 55 years old, different only in their training state, regarding the reaction of the resonance frequence. It would be established, that the resonance frequence of the A. radials in non-trained is more than the double as in trained probands. The middle resonance frequence increases continuous with the age in both groups. A high resonance frequence in thong-minded trained was not observed, only one time a normal, 63 times a reduced arterial resonance frequence. Under the condition, that the arterial resonance frequence can be regarded as an arterial index, the differences between the resonance frequence of both groups mean, that the biological age of the arteries in trained is on an average younger as in non-trained.